微生物学报

Species	Lignin (%)	Cinnamyl alcohol precursor (%)
		p-coumaryl alcohol	Coniferyl alcohol	Sinapyl alcohol
Hardwood	25−38	0−8	25−50	45−75
Softwood	20−30	< 5	> 95	0
Herbaceous plant	8−15	5−35	35−80	20−55

Species	Lignin (%)	Cinnamyl alcohol precursor (%)
		p-coumaryl alcohol	Coniferyl alcohol	Sinapyl alcohol
Hardwood	25−38	0−8	25−50	45−75
Softwood	20−30	< 5	> 95	0
Herbaceous plant	8−15	5−35	35−80	20−55

Lignin-degrading strains	Growth characteristics	Source	References
Aneurinibacillus sp. LD3	Degradation of lignin up to 61.28% at 50 ℃	Soil samples covered with sawdust	[21]
Bacillus amyloliquefaciens SL-7	The strain can be composted for 13 days at temperatures between 55 ℃ and 62 ℃	Tobacco straw	[39]
Bacillus amyloliquefaciens HZ11	Degradation of lignin up to 46.7% at 50 ℃	Fungus chaff, straw and cow dung composted to produce seedling substrate	[40]
Serratia sp. AXJ-M	Strain was able to survive in heat stress environments ranging from 50−80 ℃, with 50 ℃ being the optimal temperature for growth	Soil	[41-42]
Stenotrophomonas maltophilia LS-1	The optimal conditions for enzyme production by the strain were pH 8.0 and temperature 15 ℃	Frozen soil	[43]
Arthrobacter sp. C2	The optimal conditions for enzyme production activity of this strain were initial pH 6.7 and temperature 14.9 ℃	Frozen soil	[22]
Enterobacter sp. YP-2023-9	The strain can tolerate at least 0.60% nicotine	Cow dung	[44]
Chaetomium sp. CS1	Promoting the release of nicotinic actives from tobacco stover	Bottomless sea sediment	[45]
Sutcliffiella sp. NC1	Alkaliphilic bacteria, which do not survive at pH 6.0, grow normally at pH 8.0−11.0	Soil	[24]
Bacillus ligniniphilus L1	The strain has an optimum growth temperature of 30 ℃, an optimum pH of 9.0 and can grow at 0−10% total salt concentration, with an optimum salt concentration of 2% (W/V)	Bottomless sea sediment	[26]
Pseudomonas putida KT2440	This strain is well adapted to salt osmotic stress, and at the same time has remarkable metabolic diversity and withstands difficult redox reactions and operational stresses	Soil	[46-47]

Lignin-degrading strains	Growth characteristics	Source	References
Aneurinibacillus sp. LD3	Degradation of lignin up to 61.28% at 50 ℃	Soil samples covered with sawdust	[21]
Bacillus amyloliquefaciens SL-7	The strain can be composted for 13 days at temperatures between 55 ℃ and 62 ℃	Tobacco straw	[39]
Bacillus amyloliquefaciens HZ11	Degradation of lignin up to 46.7% at 50 ℃	Fungus chaff, straw and cow dung composted to produce seedling substrate	[40]
Serratia sp. AXJ-M	Strain was able to survive in heat stress environments ranging from 50−80 ℃, with 50 ℃ being the optimal temperature for growth	Soil	[41-42]
Stenotrophomonas maltophilia LS-1	The optimal conditions for enzyme production by the strain were pH 8.0 and temperature 15 ℃	Frozen soil	[43]
Arthrobacter sp. C2	The optimal conditions for enzyme production activity of this strain were initial pH 6.7 and temperature 14.9 ℃	Frozen soil	[22]
Enterobacter sp. YP-2023-9	The strain can tolerate at least 0.60% nicotine	Cow dung	[44]
Chaetomium sp. CS1	Promoting the release of nicotinic actives from tobacco stover	Bottomless sea sediment	[45]
Sutcliffiella sp. NC1	Alkaliphilic bacteria, which do not survive at pH 6.0, grow normally at pH 8.0−11.0	Soil	[24]
Bacillus ligniniphilus L1	The strain has an optimum growth temperature of 30 ℃, an optimum pH of 9.0 and can grow at 0−10% total salt concentration, with an optimum salt concentration of 2% (W/V)	Bottomless sea sediment	[26]
Pseudomonas putida KT2440	This strain is well adapted to salt osmotic stress, and at the same time has remarkable metabolic diversity and withstands difficult redox reactions and operational stresses	Soil	[46-47]

极端微生物降解木质素的研究进展

PDF下载

孙梦瑶 ^# , 常潇 ^# , 康志超 , 于洪文 ^*

微生物学报 | 综述 2024,64(11): 4036-4053

收起

微生物学报 | 综述 2024, 64(11): 4036-4053

极端微生物降解木质素的研究进展

全屏

孙梦瑶^#, 常潇^#, 康志超, 于洪文^*

作者信息

中国科学院东北地理与农业生态研究所, 黑土地保护与利用全国重点实验室, 湿地生态与环境重点实验室, 吉林长春 130102

Lignin degradation by extremophiles

Mengyao SUN^#, Xiao CHANG^#, Zhichao KANG, Hongwen YU^*

Affiliations

State Key Laboratory of Black Soils Conservation and Utilization, Key Laboratory of Wetland Ecology and Environment, Northeast Institute of Geography and Agroecology, Chinese Academy of Sciences, Changchun 130102, Jilin, China

出版时间: 2024-07-24 doi: 10.13343/j.cnki.wsxb.20240261

文章导航

摘要

收起

木质素是自然界最丰富的芳香族生物聚合物资源，由于其复杂性和高度聚合的芳香族结构，木质素很难被普通微生物降解。来自极端环境的木质素降解微生物被认为是木质素生物加工的合适候选者。本文对几类能够降解木质素的极端微生物及其产生的极端酶进行了归纳总结，并阐明了极端酶的性质、催化机制及其代谢途径。讨论了通过多组学方法鉴定新型极端微生物和极端酶的前景，并对极端微生物的开发和利用方法进行了展望，以期为后续筛选和开发更加高效的木质素降解菌株提供参考。

关键词

木质素 / 极端微生物 / 极端酶 / 生物降解 / 组学技术

Abstract

收起

Lignin, the most abundant aromatic biopolymer resource in the nature, is difficult to be degraded by common microorganisms due to its complexity and highly aggregated aromatic structure. Lignin-degrading microorganisms from extreme environments are considered as suitable candidates for lignin bioprocessing. This review summarizes several types of extremophiles capable of degrading lignin and the extremozymes produced by them and elucidates the properties, catalytic mechanisms, and metabolic pathways of the extremozymes. Furthermore, this article discusses the prospects for the identification of novel extremophiles and extremozymes by multi-omics and makes an outlook on the development and utilization methods of extremophiles, with a view to providing a reference for the subsequent screening and development of more efficient lignin-degrading strains.

Key words

lignin / extremophiles / extremozymes / biodegradation / omics

引用本文

孙梦瑶, 常潇, 康志超, 于洪文. 极端微生物降解木质素的研究进展. 微生物学报, 2024 , 64 (11) : 4036 -4053 . DOI: 10.13343/j.cnki.wsxb.20240261

Mengyao SUN, Xiao CHANG, Zhichao KANG, Hongwen YU. Lignin degradation by extremophiles[J]. Acta Microbiologica Sinica, 2024 , 64 (11) : 4036 -4053 . DOI: 10.13343/j.cnki.wsxb.20240261

正文

收起

木质纤维素生物质是地球上最丰富的自然资源，其中木质素是主要成分之一，是芳香族聚合物的重要资源^[1]。由于木质素结构的异质性和复杂性，在其解聚和降解的过程中，往往需要涉及与物理、化学和生物相关的多种方法，如电子束辐照改善木质素的降解性能^[2]、使用离子液体改性木质素^[3]、利用漆酶(laccase, Lac)、木质素过氧化物酶(lignin peroxidase, LiP)、锰过氧化物酶(manganese peroxidase, Mnp)、染料脱色过氧化物酶(dye decolorization peroxidase, DyP)等进行酶促降解^[4]。木质素是一种普遍存在的碳能源，寻找合适的方法(尤其是绿色环保且依赖于自然界微生物的方法)降解木质素以增强其应用价值是非常有意义的。极端微生物是最适合生活在极端环境中的微生物的总称，根据其生存极端环境的不同，可分为嗜冷微生物、嗜热微生物、嗜酸微生物、嗜碱微生物、嗜盐微生物、嗜旱微生物、嗜压微生物等，特殊的生存条件导致其具有特殊的遗传背景和代谢途径，并可产生功能特殊的酶类和活性物质^[5]。

在木质纤维素的应用中，木质素的降解十分关键。在农业领域，全球每年都会产生大量的含木质素的固体废弃物，如秸秆、甘蔗渣、糠壳^[6]，而只有少数得到了有效的再利用，大部分被直接丢弃或焚烧^[7]，尤其在寒冷地区，这些含木质素废物的降解和转化效率更低^[8]。在畜牧业中，植物性饲料中木质素的存在降低了动物饲料的消化利用率，影响畜牧业的健康和快速发展^[9-10]。在工业领域，由于木质素难以降解，阻碍了生物质资源转化为生物燃料、生物塑料、生物肥料和其他增值化学品^[11]。因此，木质素的高效降解是控制环境污染、提高畜牧业饲料利用率和经济效益、实现木质素资源化利用的有效途径，而开发能够耐受木质素生物精炼过程中的高温、低温、高渗透压、强酸、强碱条件的酶或微生物至关重要。极端微生物已经进化出许多与代谢活动、生存和保护策略有关的适应机制，使其能够适应高海拔地区、深海、热液、高盐度和极地等极端环境并大量繁殖，产生不同种类的活性酶。由于其独特的生理特性和酶学特征，极端微生物和极端酶从生产医疗和工业用途的生物分子到对废水和沉积物中的有毒污染物进行生物修复均具有潜在的应用价值^[12-14]。本文综述了极端微生物降解木质素的策略，介绍了降解木质素的极端酶及其降解机制，讨论了通过多组学方法鉴定新型极端微生物和极端酶的前景，并对极端微生物的开发和利用方法进行了展望。如果能够获得高效降解木质素的极端微生物，并深入了解其降解机制，就能够在极端的生物过程中降解/发酵木质素，促进该生物资源的利用。

1 木质素的结构和组成

收起

木质素是一种异质性多酚材料，主要由3种肉桂醇前体(单木质素)组成：芥子醇(3, 5-dimethoxy-4-hydroxyphenylpropane, S unit)、松柏醇(4-hydroxy-3-methoxyphenylpropane, G unit)和对香豆醇(4-hydroxyphenylpropane, H unit) (图1A)^[1]。天然木质素的组成和结构随其物种的不同而变化，并表现出局部和季节的变化。天然木质素通常可分为3种类型：硬木(被子植物)、软木(裸子植物)和草本(禾本科)木质素。硬木木质素主要由松柏醇和芥子醇前体组成，软木木质素主要由松柏醇前体组成，草本木质素具有3种类型的前体^[15]。表1显示了不同木质纤维素生物质中木质素和3种肉桂醇前体的含量^{[1, 15]}。

在生物聚合(木质化)过程中，这些木质素单体通过氧化缩合连接在一起，形成多种键^[16] (图1B)。自然界中的木质素聚合不受酶或蛋白质控制，一旦产生自由基，就是一个纯粹的化学过程，可分为2个阶段：在第一阶段，酚类木质素单体自由基二聚形成β-O-4、β-β和β-5脱氢二聚体，这是木质素聚合物中最常见的键；随着聚合的进行，低聚酚末端单元通过5-5和4-O-5型键连接；同时，在木质素单体和预先形成的β-芳基醚二聚末端单元之间形成β-1型连接；这种自由基偶联过程在木质素结构中缺乏规则和有序的重复单元(图1C)，因此木质素的利用受到限制^{[1, 17]}。

2 降解木质素的极端微生物

收起

生物转化和降解是利用木质素的最重要机制之一。目前已知许多真菌和细菌均具有优异的木质素降解能力，它们对木质素的分解和代谢具有多种不同策略，并且不产生二次污染，绿色环保。研究表明热红短芽孢杆菌(Brevibacillus thermoruber)^[18]、褐色喜热裂孢菌(Thermobifida fusca)^[19]、热解纤维梭菌(Clostridium thermocellum)^[20]、解硫胺素芽孢杆菌(Aneurinibacillus sp.) LD3^[21]、节杆菌(Arthrobacter sp.) C2^[22]等极端微生物均具有降解木质素的作用。近年来，许多研究倾向于筛选并构建出单一高效的木质素降解菌株或构建人工复合菌系协同降解木质素，以提高其利用价值。

2.1 单一菌株降解木质素

研究表明，木质素在碱性溶液中的溶解度随所用溶剂的极性、溶液黏度和所用碱性试剂的阳离子半径而产生较大变化，高pH值不仅有助于提高木质素的溶解度，而且还能降低木质素粒径^[23]。为了在碱性溶液中实现木质素生物转化，Xu等^[24]在pH 11.0的碱性条件下筛选出了几种耐碱木质素分解细菌，其中萨克利夫氏菌(Sutcliffiella sp.) NC1被证明是一种嗜碱细菌，其木质素过氧化物酶在碱性条件下表现出高度活性。Jiang等^[22]为解决中国寒冷地区大部分富含木质纤维素的农业剩余物降解受限的问题，从−10 ℃的冻土中分离出一株耐冷细菌节杆菌C2，该菌株产酶活性的最佳条件为初始pH 6.7、温度14.9 ℃。研究发现耐冷菌株C2可应用于造纸黑液的处理，该菌株具有良好的冷适应性和木质素降解能力，在15 ℃时，3 g/L的木质素溶液的降解率为65.5%，矿化率为43.9%^[25]。此外，Wu等^[21]筛选出了耐热碱木质素降解菌株解硫胺素芽孢杆菌LD3并将其应用于餐厨垃圾堆肥过程，该菌株具有较强的碱木质素降解能力和木质素分解酶活性，在降解过程中产生许多低分子量化合物，促进了芳香结构的形成。同时，由于木质素降解过程中会产生芳香族化合物，通常对细胞有毒，所以木质素降解过程需要微生物能够耐受这些芳香族化合物的抑制作用或毒性。因此，为了克服酚类抑制剂过量的问题，需要利用能够分泌耐热木质素酶并降解这些酚类化合物的微生物。从深海沉积物中分离得到的耐碱嗜盐细菌嗜木质素芽孢杆菌(Bacillus ligniniphilus) L1表现出显著的木质素降解能力，其木质素解聚混合物中含有数十种芳香族化合物^[26]。此外，韩宇杰等^[27]从森林土壤、秸秆还田土及腐烂的木材等样品中筛选出了3株靶向降解木质素的细菌菌株，在pH5.0−9.0，培养温度10−50 ℃下，3株细菌均可以正常生长，并具有降解木质素的能力。Kang等^[28-29]在吉林大安国家农田生态系统野外观测研究站玉米田土壤中筛选出一株耐冷类节杆菌(Paenarthrobacter sp.) KN0901，该菌株在15 ℃下可同时降解莠去津和秸秆复合污染物，并且莠去津与秸秆共同降解使该菌株的生化反应水平有所提高，进一步消除了莠去津对植物的毒性。综上所述，使用单一菌株降解木质素时，可以进行定向筛选，从而得到适应性更好的微生物，以提高木质素降解率；同时，也可以考虑使用单一菌株对含木质素的废弃物和其他化学污染物共同降解，这可能会促进微生物的降解作用。

2.2 多种微生物协同降解木质素

由于木质素结构的异质性和生物转化途径的可变性，单一菌株的木质素解聚效率有限，高效的微生物反应需要一套复杂的木质素水解酶，而单一微生物无法产生^[30]。构建微生物共培养体系或自然富集培养细菌群落已被认为是提高木质素分解效率的方法，微生物联合体可以克服单一菌株降解效率受限的问题，整合多种微生物的木质素代谢途径，形成更全面的木质素代谢网络^[31-32]。微生物群落，特别是嗜极微生物群落的发现，对于制定实现高效木质素生物降解过程的策略至关重要。例如，Wang等^[33]富集了一组高温(50 ℃)木质素降解菌群，并优化了木质素降解的培养条件，结合高通量测序技术，分析了复合菌群的协同效应，并将高温木质素降解菌群处理过的稻草用于纤维膜的制备。Yadav等^[34]从纸浆造纸厂废水污染场地中分离出枯草芽孢杆菌(Bacillus subtilis)、生癌肠杆菌(Enterobacter cancerogenus)和地衣芽孢杆菌(Bacillus licheniformis)，在共同降解木质素的过程中能够产生高分子量的群体感应信号分子(N-acyl homoserine lactone, AHL)，AHL的产生导致生物膜的形成，形成生物膜的菌群木质素降解率比单一培养的枯草芽孢杆菌、癌肠杆菌和地衣芽孢杆菌高出34%−39%；同时，将联合菌群用于废水处理后，水中的主要有机污染物减少，从而证实了活性生物转化和木质素的生物降解。为探索低温下加速玉米秸秆分解和回归的有效途径，Cheng等^[35]以生物炭为载体，利用9株耐冷纤维素、半纤维素和木质素降解菌株，制备了新型耐冷秸秆降解菌群；在10−15 ℃下，该耐冷菌群处理50 d后玉米秸秆最大降解率达到59.3%，并增加了土壤中有机质、磷、钾的含量；同时，该菌群的添加提高了土壤中微生物群落多样性，改变了特异性选择，使节杆菌(Arthrobacter)、假单胞菌(Pseudomonas)和泛菌(Pantoea)成为土壤微生物中的优势菌属，促进了玉米秸秆的降解。在我们之前的研究中，通过土壤富集获得了由鞘氨醇杆菌(Sphingobacterium)、黄杆菌(Flavobacterium)、地杆菌(Pedobacter)、节杆菌(Arthrobacter)、代夫特菌(Delftia)和鞘氨醇单胞菌(Spingomonas)组成的耐低温木质纤维素降解菌群，该菌群在15 ℃的低温下对废纸和秸秆的降解率达到了21.8%和52.0%^[36]。此外，能降解莠去津和秸秆复合污染物的KN0901可以和另一株红球菌(Rhodococcus sp.) KD0901构成人工微生物群落KDN0901，该人工复合群落对不同pH和温度条件(分别为5.0−9.0和4−35 ℃)表现出高度适应性，尤其是在低温下可同时降解莠去津和邻苯二甲酸二丁酯^[37]。综上所述，从各种极端环境中筛选具有显著木质素降解能力的极端微生物，构建能够协同降解木质素的微生物菌群，也是提高微生物降解木质素效率的重要途径。此外，复合极端微生物群落的降解能力往往不是单一的，在多种微生物共同作用下，包括木质纤维素在内的多种固体废弃物均能在一定程度上得到降解和转化^[38] (已报道的能够降解木质素的极端微生物的详细信息见表2)。

3 降解木质素的极端酶及其降解机制

收起

3.1 Lac、Lip、MnP和DyP

Lac (EC: 1.10.3.2)是一种含有4个铜离子的多酚氧化酶，属于铜蓝氧化酶，以单体糖蛋白的形式存在，它存在于蘑菇、细菌和植物以及空气中，催化酚类和非酚类化合物的氧化，其中耐热Lac是一种优异的木质素降解绿色催化剂，其副产品无化学污染，需求广泛^[48]。Khan等^[49]从高地芽孢杆菌(Bacillus altitudinis) SL7中克隆了具有类似Lac活性的CotA蛋白编码基因，并在大肠杆菌中进行了表达，纯化的蛋白酶CotA-SL7在宽泛的温度(30−90 ℃)和pH值(3.0−10.0)范围内都具有活性，在55 ℃和pH 5.0时活性最佳，该研究还证明了CotA-SL7能裂解木质素聚合物并打开苯环，具有较高的催化效率。此外，从嗜木质素芽孢杆菌(Bacillus ligniniphilus) L1中获得的稳定性Lac也表现出优异的木质素降解能力，碱性木质素和磨碎木质素的降解率分别达到67.0%和80.9%^[50]。

LiP (EC: 1.11.1.14)是一种带有血红素蛋白的糖基化酶，使用过氧化氢作为氧受体催化木质素聚合物中存在的不同键的裂解。与Lac相比，LiP由于其高氧化还原电位，无需介体即可有效氧化酚类和非酚类木质素结构^[51]。LiP可以氧化木质素的重要键，例如β-O-4和C−C，这表明它在木质素降解和相关工业应用方面有巨大的潜力^[52]。Rath等^[53]从印度的一个生物保护区的土壤样品中分离和筛选出一株产LiP的菌株，通过响应面分析发现LiP在pH 10.5、温度30 ℃、底物浓度0.115 mmol/L和培养时间42 h时具有最大活性。

MnP (EC: 1.11.1.13)是含血红素的过氧化物酶之一，主要由寄生在木材上的白腐菌和担子菌产生，在木质素的初始解聚中发挥重要作用，凭借其较高的氧化还原电位，MnP能够促进酚类木质素的显著降解，并最终释放二氧化碳^[54]。从白腐真菌(Irpexlacteus sp.) CD2中纯化的耐热MnP对金属离子和有机溶剂表现出显著的耐受性，并且在3.5−6.0的pH范围内高度稳定，最佳温度为70 ℃^[55]。Zhang等^[47]研究表明，MnP通过诱导杨树预水解液中的木质素聚合来提高木质素的去除率，同时保证糖类物质含量不受影响。当Lac、LiP和MnP联合使用降解碱性木质素时，碱性木质素的降解率最高，但其结构完整性较低^[56]。

DyP (EC: 1.11.1.19)是一类血红素过氧化物酶，普遍存在于细菌、放线菌、真菌、古细菌和后生动物中，其在系统发育、结构和机制上与其他过氧化物酶(LiP和MnP)不同^[57]。Yu等^[58]研究发现，嗜热放线菌(Saccharomonospora viridis)可以产生稳定的耐热DyP，其在较宽的pH范围(5.0−10.0)和较高温度(50−80 ℃)下均有活性。

传统的木质素酶在较窄的反应条件范围内是合适的催化剂，但在高温和极端pH等条件下则表现出较低的稳定性^[59]。因此，极端酶是恶劣条件下木质素降解的关键突破口。然而，截至目前极端酶尚未在木质素生物精炼中大规模应用，阻碍极端酶广泛应用的原因之一，是通过非常规分子生物学技术难以高效分离和培养在异源系统中高表达的极端酶^[60]。在今后的研究中应当致力于开发分离和培养极端酶的生物工具，以实现木质素的高效利用。

3.2 降解机制

如图2A所示，木质素衍生的芳香族化合物的降解至少涉及5种不同的途径^[61-67]。(1) β-芳基醚降解途径，木质素降解菌株首先将愈创木基甘油-β-愈创木基醚(guaiacylglycerol-β-guaiacyl ether, GGE)转化为α-(2-甲基苯氧基)-β-羟基丙香草酮[α-(2-met hoxyphenoxy)-β-hydroxypropiovanillone, MPHPV]，随后MPHPV的醚键通过还原裂解产生β-羟基丙吡香草酮(β-hydroxyproppiovanillone, HPV)和愈创木酚。(2) 联苯降解途径，5, 5-脱氢二香草酸(5, 5-dehydrodivanillate, DDVA)首先通过木质素降解菌株进行O-去甲基化，产生2, 2′, 3-三羟基-3′-甲氧基-5, 5′-二羧基联苯(2, 2′, 3-trihydroxy-3′-methoxy-5, 5′-dicarboxybiphenyl, OH-DDVA)，OH-DDVA在双加氧酶的作用下降解为4, 11-二羧基-8-羟基-9-甲氧基-2-羟基-6-氧代-6-苯基六-2, 4-二烯酸酯(4, 11-dicarboxy-8-hydroxy-9-methoxy-2-hydroxy-6-oxo-6-phenylhexa-2, 4-dienoate, DCHM-HOPDA)，而后生成香草酸(vanillate, VAN)；VAN代谢是通过O-去甲基化生成原儿茶酸(protocatechuate, PCA)来解释的；PCA的降解取决于PCA 4, 5-裂解途径。在该途径中，PCA的降解可通过4, 5-裂解途径分为3种不同的代谢途径，在最典型的分支之一中，PCA最初转化为4-羧基-2-羟基粘康酸酯-6-半醛(4-carboxy-2-hydroxymuconate-6-semialdehyde, CHMS)；CHMS无需酶促作用即可转化为分子内半缩醛形式，然后氧化为2-吡喃酮-4, 6-二羧酸酯(2-pyrone-4, 6-dicarboxylate, PDC)；接着PDC生成酮和烯醇形式的4-草酰中康酸酯(4-oxalomesaconate, OMA)，最终生成4-羧基-4-羟基-2-氧代己二酸酯(4-carboxy-4-hydroxy-2-oxoadipate, CHA)，进入三羧酸循环(tricarboxylic acid cycle, TCA)。(3) 阿魏酸途径，阿魏酸最初通过生物转化转化为VAN；而后转化为PCA，进入PCA降解途径。(4) 对香豆酸降解途径，对香豆酸(4-hydroxycinnamic acid, HCNA)转化为4-羟基苯甲酸钠，随后进入PCA降解。(5) 丁香醛降解途径，丁香醛(syringaldehyde, SA)首先转化为丁香酸盐，随后在丁香酸脱甲基酶的作用下转化为3-O-没食子酸甲酯(3-O-methylgallate, 3MGA)，化合物3MGA转化为没食子酸盐，观察到的肉桂酸与对羟基苯基单元的氧化有关，这些是芥子基紫丁香基单元的基本构建单元，芥子基单元是木质素结构的一部分。

木质素降解的第一步通常由外源性氧化还原酶如Lac、LiP、MnP、DyP完成；随后，产生的异质混合物的芳香族产物通过各种胞内酶进行代谢，包括双加氧酶、脱羧酶、甲基转移酶、水解酶、脱氢酶、超氧化物歧化酶和谷胱甘肽转移酶(与这些酶相关的基因在环状基因组中的表达和定位如图2B所示)，这些胞内酶将芳香族产物转化为中心中间体；最后，中间体通过β-酮戊二酸途径进行环裂解并进一步分解进入中心碳代谢；其中，最重要的机制是通过上游途径产生中间产物，如香草醛、苯甲酸和原儿茶酸等，随后，中间产物通过双加氧酶环裂解反应进入下游途径，最终进入TCA途径^{[63-65, 67]}。

4 多组学技术筛选极端微生物降解木质素

收起

经典的依赖适宜条件培养的方法只能识别某一环境中10%−20%的微生物群落。相比之下，包括宏基因组学、宏转录组学、宏蛋白质组学和代谢组学在内的无需对微生物进行分离培养的现代技术，使得我们能够对不同环境中的微生物群落的木质素降解机制进行探究^[68]。宏基因组学，也称为微生物环境基因组学，需要从环境样本中发现的整个微生物种群中直接提取DNA。此后，构建宏基因组文库并使用基因组研究策略来研究这些微生物的遗传组成和群落功能。宏基因组学于1998年首次提出，最初用于土壤和海洋微生物的分类和鉴定，后来推广用于其他环境，有助于发现新的微生物物种和遗传元素^[69-70]。宏转录组学是对微生物RNA的综合分析，以研究特定时间间隔内的全基因组转录活性和调控机制^[71]。与宏基因组学不同，宏转录组学可以识别代谢活跃的微生物群体，并揭示微生物生长过程中的通用基因表达模式^[72]。宏蛋白质组学是特定时间点环境中微生物组的总蛋白质补充。虽然宏基因组学可以揭示微生物群落中潜在基因的功能，宏转录组学可以深入了解基因表达和活性，但其细胞定位和活性调节发生在蛋白质水平，因此从转录组和蛋白质组获得的信息会有所不同^[73]，蛋白质组学是一种用于表征细胞、组织或生物体中存在的蛋白质并生成蛋白质表达定量图谱的方法。蛋白质组学分析可以揭示生物体的实际表达谱，这是使用纯基因组方法无法实现的^[74]。代谢组学是一种基于核磁共振(nuclear magnetic resonance, NMR)、气相色谱-质谱(GC-MS)、液相色谱-质谱(liquid chromatography-mass spectrometry, LC-MS)、毛细管电泳-质谱(capillary electrophoresis-mass spectrometry, CE-MS)的新兴技术^[75]。代谢组学分析是对生物系统中初级和次级代谢物进行检测、分类和定量的完整过程，使得复杂基因和蛋白质网络的研究变得简单直观，代谢组分析是一种强大的工具，可以帮助确定微生物降解木质素过程中产生的小分子的种类和丰度，与基因和蛋白质相比，代谢物的产生是生物体中最下游的活动^[76]。使用单一方法来调查高度复杂的微生物群落将导致描述不完整。为了克服这个问题，许多研究人员正在利用整合宏基因组学、宏转录组学、宏蛋白质组学和代谢组学的多组学方法来研究某一环境中的微生物群落(各组学之间的关系如图3所示)。结合宏基因组和宏蛋白质组分析，可以描述在精确条件下共同生长过程中产生的关键酶，而不仅仅是识别微生物群落的遗传信息。此外，宏基因组学和宏转录组学能够预测微生物群的功能潜力，而无需完整的基因组重建。总而言之，通过整合这些不同层面的数据，可以更全面地理解微生物群落的结构、功能，从而促进微生物的相关应用。

Hassa等^[77]通过整合宏基因组测序、宏转录组学、宏蛋白组学和代谢组学方法，对沼气发酵中的微生物群落进行分析，深入了解了其微生物群落的组成、性能、群落成员之间的相互关系，这为厌氧发酵条件下木质纤维素降解菌的筛选奠定了基础。Tang等^[78]的研究发现，竹节虫(Cyrtotrachelus buqueti)的肠道分泌蛋白的纤维素降解率为30.23%−38.24%，半纤维素为58.56%−70.96%，木质素为28.71%−43.43%；而微生物群的纤维素降解率为28.41%−38.31%，半纤维素为53.45%−66.15%，木质素为25.65%−38.37%；该研究综合宏基因组测序和转录组测序分析确定了核心微生物群落和碳水化合物活性酶(carbohydrate-active enzyme, CAZymes)，显示出微生物组和转录组的密切相关性，揭示了竹节虫肠道微生物降解木质纤维素的机制。此外，还可以利用宏基因组测序技术分析已有的耐低温秸秆降解菌群之间的相关性，对其进行优化，降低菌群的复杂性，以增强实际应用中的优势^[79]。Wang等^[80]基于全基因组分析的宏基因组测序解释了玉米秸秆和猪粪共消化和单一消化中微生物群落的分类和功能模式，发现耐热梭状芽孢杆菌(Clostridium stercorarium)是玉米秸秆降解的特异菌种，甲烷鬃毛状菌(Methanosaeta concilii)是主要的产甲烷菌种。D’haeseleer等^[81]基于蛋白质组学和宏基因组测序技术分析了能够解构柳枝稷[Panicum virgatum (可作为生物燃料)]的嗜热微生物群，检测到了许多种类的木质素降解酶，并证实了类芽孢杆菌(Paenibacillus)在生物质解构中发挥着核心作用。Martien和Amador-Noguez^[82]指出代谢组学分析可以揭示目标基因，表征木质素等微生物燃料的代谢过程，从而实现木质素原料的利用，并提高工程微生物的应激耐受性。

由于极端微生物及其酶的特殊生理特征以及缺乏定制的分子生物学工具，研究这些微生物比普通微生物更具挑战性。基于宏基因组学、转录组学、蛋白质组学和代谢组学的微生物鉴定方法，打破了依赖于微生物分离培养方法的严格界限，使得能够对由不可培养和可培养微生物组成的任何环境样本进行深入研究。此类技术允许探测和分析大量数据，这些数据代表任何生物系统在DNA、RNA、蛋白质和代谢物水平上的整体功能和结构^[83]。综上所述，组学技术为筛选和鉴定降解木质素的极端微生物和极端酶提供了新的方向。

5 小结与展望

收起

木质素资源丰富，但结构复杂，由酚丙烷单元组成，通过不同的键合方式连接，导致其降解效率受限。近年来利用高通量测序技术以及多组学技术已经筛选出了许多木质素降解菌，并对其降解机制有了一定的了解。然而，木质素的工业应用往往需要特定的环境条件，如高温高压、极端pH等。极端微生物在恶劣的条件下仍能高效地分泌极端酶来特异性地断裂这些键合，从而降解木质素。因此，对极端微生物的开发和利用是在未来的研究中需要加强的，具体应侧重以下方面。

(1) 丰富极端微生物资源库。应当继续从不同的环境中筛选能够降解木质素的极端微生物，如冻土、热泉、深海以及食草动物的胃肠道等。同时，将筛选得到的微生物进行适应性进化，在实验室模拟极端微生物在自然环境中的进化过程，筛选出具有更强适应性和生产性能的菌株^[84]。

(2) 构建降解木质素的人工复合菌群。单一微生物菌株往往难以完全降解木质素，通过构建复合菌群，可以将分泌不同类型木质素降解酶的微生物组合在一起，这些酶可以协同作用，打破木质素的不同化学键，从而提高降解效率。复合菌群中的微生物可能具有不同的环境适应性，这样的组合可以使复合菌群在更广泛的环境条件下稳定工作，增强了木质素降解过程的稳健性；不同的微生物可能对木质素的不同部分有不同的降解偏好。通过合理组合，可以针对特定的木质素结构设计复合菌群，提高对目标化合物的选择性降解，减少不必要的副产物生成。相比于化学法，生物法降解木质素通常更为环保且成本较低；通过优化复合菌群的构建，可以减少对化学助剂的依赖，降低生产成本，同时减少化学处理可能带来的环境污染。

(3) 对筛选出的极端微生物和极端酶进行生物技术修饰。基因工程改造是通过基因编辑技术，如CRISPR/Cas9系统，可以对极端微生物的基因组进行精确的修饰，增强其在特定环境下的适应性或提高极端酶的催化效率^[85]。蛋白质工程是利用理性设计或定向进化的方法，可以改变极端酶的氨基酸序列，从而改善其热稳定性、pH耐受性或底物特异性^[86]。代谢工程是通过增加碳源前体、敲除旁路代谢途径、减少副产物生成、提高目标产物的转运等策略，优化极端微生物的代谢途径，提高其生产效率和产物的产率^[87]。微生物表面显示技术是将极端酶或其编码基因整合到微生物细胞表面蛋白上，使酶直接展示在细胞表面，这样可以在不分离酶的情况下进行催化反应，简化了反应过程并提高了酶的稳定性^[88]。

(4) 应大规模对木质素分解极端微生物进行全基因组测序，然后进行比较基因组分析和基因组挖掘，以确定木质素降解和生物漏斗路径、转录调节因子和限速机制。此外，注重木质素分解极端微生物的转录组学、蛋白质组学和代谢组学综合分析，揭示木质素的生物降解功能和机制。

基金

收起

吉林省与中国科学院科技合作高技术产业化专项基金(2023SYHZ0008)

参考文献

收起

文献

收起

参考文献引证文献

排序方式：

[1]

LIU J, LI X, LI M, ZHENG Y. Lignin biorefinery: lignin source, isolation, characterization, and bioconversion[M]//Advances in Bioenergy. Amsterdam: Elsevier, 2022: 211-270.

[2]

KUMAR A, TUMU VR, RAY CHOWDHURY S, SVS RR. A green physical approach to compatibilize a bio-based poly(lactic acid)/lignin blend for better mechanical, thermal and degradation properties[J]. International Journal of Biological Macromolecules, 2019, 121:588-600.

https://doi.org/10.1016/j.ijbiomac.2018.10.057

[3]

SZALATY TJ, KLAPISZEWSKI Ł, JESIONOWSKI T. Recent developments in modification of lignin using ionic liquids for the fabrication of advanced materials: a review[J]. Journal of Molecular Liquids, 2020, 301:112417.

https://doi.org/10.1016/j.molliq.2019.112417

[4]

ZHANG ST, XIAO JL, WANG G, CHEN G. Enzymatic hydrolysis of lignin by ligninolytic enzymes and analysis of the hydrolyzed lignin products[J]. Bioresource Technology, 2020, 304:122975.

https://doi.org/10.1016/j.biortech.2020.122975

[5]

庄滢潭, 刘芮存, 陈雨露, 杨岑玥, 余岩, 王涛, 王友亮, 宋亚军, 滕越. 极端微生物及其应用研究进展[J]. 中国科学(生命科学), 2022, 52 (2):204-222.

ZHUANG YT, LIU RC, CHEN YL, YANG CY, YU Y, WANG T, WANG YL, SONG YJ, TENG Y. Extremophiles and their applications[J]. Scientia Sinica (Vitae), 2022, 52 (2):204-222.

[6]

KESHAV PK, BANOTH C, KETHAVATH SN, BHUKYA B. Lignocellulosic ethanol production from cotton stalk: an overview on pretreatment, saccharification and fermentation methods for improved bioconversion process[J]. Biomass Conversion and Biorefinery, 2023, 13 (6):4477-4493.

https://doi.org/10.1007/s13399-021-01468-z

[7]

MU BS, TANG W, LIU T, HAO XL, WANG QW, OU RX. Comparative study of high-density polyethylene-based biocomposites reinforced with various agricultural residue fibers[J]. Industrial Crops and Products, 2021, 172:114053.

https://doi.org/10.1016/j.indcrop.2021.114053

[8]

韩宇杰, 常潇, 向海, 钟荣珍. 秸秆低温降解的研究进展[J]. 动物营养学报, 2023, 35 (6):3528-3536.

HAN YJ, CHANG X, XIANG H, ZHONG RZ. Research progress of straw degradation at low temperature[J]. Chinese Journal of Animal Nutrition, 2023, 35 (6):3528-3536.

[9]

ATUHAIRE AM, KABI F, OKELLO S, MUGERWA S, EBONG C. Optimizing bio-physical conditions and pre-treatment options for breaking lignin barrier of maize stover feed using white rot fungi[J]. Animal Nutrition, 2016, 2 (4):361-369.

https://doi.org/10.1016/j.aninu.2016.08.009

[10]

常潇, 房义, 向海, 贺志雄, 吴端钦, 钟荣珍. 不同长度秸秆对妊娠后期母羊生理代谢及其子代羔羊生长性能的影响[J]. 动物营养学报, 2021, 33 (9):5108-5118.

CHANG X, FANG Y, XIANG H, HE ZX, WU DQ, ZHONG RZ. Effects of different lengths of straws on late gestation ewe's physiological metabolism and growth performance of their offspring lambs[J]. Chinese Journal of Animal Nutrition, 2021, 33 (9):5108-5118.

[11]

XU R, ZHANG K, LIU P, HAN HW, ZHAO S, KAKADE A, KHAN A, DU DL, LI XK. Lignin depolymerization and utilization by bacteria[J]. Bioresource Technology, 2018, 269:557-566.

https://doi.org/10.1016/j.biortech.2018.08.118

[12]

SINGH P, JAIN K, DESAI C, TIWARI O, MADAMWAR D. Microbial community dynamics of extremophiles/extreme environment[M]//Microbial Diversity in the Genomic Era. Amsterdam: Elsevier, 2019: 323-332.

[13]

ZHU DC, QARIA MA, ZHU B, SUN JZ, YANG B. Extremophiles and extremozymes in lignin bioprocessing[J]. Renewable and Sustainable Energy Reviews, 2022, 157:112069.

https://doi.org/10.1016/j.rser.2021.112069

[14]

RADDADI N, CHERIF A, DAFFONCHIO D, NEIFAR M, FAVA F. Biotechnological applications of extremophiles, extremozymes and extremolytes[J]. Applied Microbiology and Biotechnology, 2015, 99 (19):7907-7913.

https://doi.org/10.1007/s00253-015-6874-9

[15]

POVEDA-GIRALDO JA, SOLARTE-TORO JC, CARDONA ALZATE CA. The potential use of lignin as a platform product in biorefineries: a review[J]. Renewable and Sustainable Energy Reviews, 2021, 138:110688.

https://doi.org/10.1016/j.rser.2020.110688

[16]

KUMAR A, ANUSHREE, KUMAR J, BHASKAR T. Utilization of lignin: a sustainable and eco-friendly approach[J]. Journal of the Energy Institute, 2020, 93 (1):235-271.

https://doi.org/10.1016/j.joei.2019.03.005

[17]

CRESTINI C, CRUCIANELLI M, ORLANDI M, SALADINO R. Oxidative strategies in lignin chemistry: a new environmental friendly approach for the functionalisation of lignin and lignocellulosic fibers[J]. Catalysis Today, 2010, 156 (1/2):8-22.

[18]

NIU JY, LI XF, QI XG, REN YP. Pathway analysis of the biodegradation of lignin by Brevibacillus thermoruber[J]. Bioresource Technology, 2021, 341:125875.

https://doi.org/10.1016/j.biortech.2021.125875

[19]

RAHMANPOUR R, REA DA, JAMSHIDI S, FÜLÖP V, BUGG TDH. Structure of Thermobifida fusca DyP-type peroxidase and activity towards Kraft lignin and lignin model compounds[J]. Archives of Biochemistry and Biophysics, 2016, 594:54-60.

https://doi.org/10.1016/j.abb.2016.02.019

[20]

AKINOSHO HO, YOO CG, DUMITRACHE A, NATZKE J, MUCHERO W, BROWN SD, RAGAUSKAS AJ. Elucidating the structural changes to Populus lignin during consolidated bioprocessing with Clostridium thermocellum[J]. ACS Sustainable Chemistry & Engineering, 2017, 5 (9):7486-7491.

[21]

WU XY, AMANZE C, WANG JS, YU ZJ, SHEN L, WU XL, LI JK, YU RL, LIU YD, ZENG WM. Isolation and characterization of a novel thermotolerant alkali lignin-degrading bacterium Aneurinibacillus sp. LD3 and its application in food waste composting[J]. Chemosphere, 2022, 307 (Pt 3):135859.

[22]

JIANG C, CHENG Y, ZANG HL, CHEN X, WANG Y, ZHANG YT, WANG JM, SHEN XH, LI CY. Biodegradation of lignin and the associated degradation pathway by psychrotrophic Arthrobacter sp. C2 from the cold region of China[J]. Cellulose, 2020, 27 (3):1423-1440.

https://doi.org/10.1007/s10570-019-02858-3

[23]

MELRO E, FILIPE A, SOUSA D, VALENTE AJM, ROMANO A, ANTUNES FE, MEDRONHO B. Dissolution of kraft lignin in alkaline solutions[J]. International Journal of Biological Macromolecules, 2020, 148:688-695.

https://doi.org/10.1016/j.ijbiomac.2020.01.153

[24]

XU ZX, LI J, LI PP, CAI CG, CHEN ST, DING BN, LIU SM, GE MS, JIN MJ. Efficient lignin biodegradation triggered by alkali-tolerant ligninolytic bacteria through improving lignin solubility in alkaline solution[J]. Journal of Bioresources and Bioproducts, 2023, 8 (4):461-477.

https://doi.org/10.1016/j.jobab.2023.09.004

[25]

WANG Y, CHEN X, WU BW, MA T, JIANG HY, MI YZ, JIANG C, ZANG HL, ZHAO XY, LI CY. Potential and mechanism for bioremediation of papermaking black liquor by a psychrotrophic lignin-degrading bacterium, Arthrobacter sp. C2[J]. Journal of Hazardous Materials, 2022, 439:129534.

https://doi.org/10.1016/j.jhazmat.2022.129534

[26]

ZHU DC, TANABE SH, XIE CX, HONDA D, SUN JZ, AI LZ. Bacillus ligniniphilus sp. nov., an alkaliphilic and halotolerant bacterium isolated from sediments of the South China Sea[J]. International Journal of Systematic and Evolutionary Microbiology, 2014, 64(Pt 5):1712-1717.

[27]

韩宇杰, 常潇, 向海, 高劲松, 钟荣珍. 秸秆木质素降解细菌的筛选及其降解条件优化[J]. 动物营养学报, 2023, 35 (7):4740-4751.

HAN YJ, CHANG X, XIANG H, GAO JS, ZHONG RZ. Isolation of straw lignin-degrading bacteria and optimization of their degradation conditions[J]. Chinese Journal of Animal Nutrition, 2023, 35 (7):4740-4751.

[28]

KANG ZC, WANG TY, HAN XR, WANG CX, ZHANG SQ, YU HW. Whole genome sequencing revealed the capability of Paenarthrobacter sp. KN0901 to simultaneously remove atrazine and corn straw at low temperatures: from gene identification to empirical validation[J]. Journal of Hazardous Materials, 2024, 471:134251.

https://doi.org/10.1016/j.jhazmat.2024.134251

[29]

KANG ZC, YANG Y, WANG CX, KANG YY, WANG TY, ZHU GP, HAN XR, YU HW. Atrazine decontamination by a newly screened psychrotroph Paenarthrobacter sp. KN0901 in an aquatic system: metabolic pathway, kinetics, and hydroponics experiment[J]. Journal of Hazardous Materials, 2023, 457:131764.

https://doi.org/10.1016/j.jhazmat.2023.131764

[30]

GU DC, XIANG XJ, WU YC, ZENG J, LIN XG. Synergy between fungi and bacteria promotes polycyclic aromatic hydrocarbon cometabolism in lignin-amended soil[J]. Journal of Hazardous Materials, 2022, 425:127958.

https://doi.org/10.1016/j.jhazmat.2021.127958

[31]

HARSHVARDHAN K, SURI M, GOSWAMI A, GOSWAMI T. Biological approach for the production of vanillin from lignocellulosic biomass (Bambusa tulda)[J]. Journal of Cleaner Production, 2017, 149:485-490.

https://doi.org/10.1016/j.jclepro.2017.02.125

[32]

JAGMANN N, PHILIPP B. Design of synthetic microbial communities for biotechnological production processes[J]. Journal of Biotechnology, 2014, 184:209-218.

https://doi.org/10.1016/j.jbiotec.2014.05.019

[33]

WANG JH, ZHANG W, WU CY, HONG YH, SHEN GN, WANG WW, TANG HZ, MOCHIDZUKI K, CUI ZJ, KHAN A, WANG WD. Synergistic analysis of lignin degrading bacterial consortium and its application in rice straw fiber film[J]. The Science of the Total Environment, 2024, 927:172386.

https://doi.org/10.1016/j.scitotenv.2024.172386

[34]

YADAV S, TRIPATHI S, PURCHASE D, CHANDRA R. Development of a biofilm-forming bacterial consortium and quorum sensing molecules for the degradation of lignin-containing organic pollutants[J]. Environmental Research, 2023, 226:115618.

https://doi.org/10.1016/j.envres.2023.115618

[35]

CHENG Y, HUANG MY, SHEN XH, JIANG C. Enhanced cornstalk decomposition by a psychrotrophic bacterial consortium comprising cellulose, hemicellulose, and lignin degraders with biochar as a carrier for carbonneutrality[J]. Bioresource Technology, 2022, 344 (Pt B):126259.

[36]

康志超. 耐低温木质纤维素降解菌群的构建及其应用研究[D]. 哈尔滨: 中国科学院大学(中国科学院东北地理与农业生态研究所)硕士学位论文, 2019.

KANG ZC. Study on the construction and application of low-temperature-tolerant lignocellulosic degradation flora[D]. Harbin: Master's Thesis of Northeast Institute of Geography and Agroecology, Chinese Academy of Sciences, 2019 (in Chinese).

[37]

KANG ZC, HAN XR, WANG CX, YU HW. Artificial microbial consortium for simultaneous removal of dibutyl phthalate and atrazine combined pollutants at low temperatures[J]. Chemical Engineering Journal, 2024, 482:149075.

https://doi.org/10.1016/j.cej.2024.149075

[38]

DUAN ZX, WANG QY, WANG TY, KONG XF, ZHU GP, QIU GK, YU HW. Application of microbial agents in organic solid waste composting: a review[J]. Journal of the Science of Food and Agriculture, 2024, 104 (10):5647-5659.

https://doi.org/10.1002/jsfa.13323

[39]

MEI JF, SHEN XB, GANG LP, XU HJ, WU FF, SHENG LQ. A novel lignin degradation bacteria: Bacillus amyloliquefaciens SL-7 used to degrade straw lignin efficiently[J]. Bioresource Technology, 2020, 310:123445.

https://doi.org/10.1016/j.biortech.2020.123445

[40]

郭玲玲, 江志阳, 陶姝宇, 于淼, 朱晓琳, 陈丽媛. 耐高温木质素降解菌筛选及降解效果研究[J]. 微生物学杂志, 2022, 42 (4):64-69.

GUO LL, JIANG ZY, TAO SY, YU M, ZHU XL, CHEN LY. Screening and degradation effect of high temperature resistant lignin degradable bacteria[J]. Journal of Microbiology, 2022, 42 (4):64-69.

[41]

汪流伟, 王燕林, 惠淑雨, 冯齐齐, 程宓丽菡, 冯瑞, 罗文怡, 李亚婷, 陈瑶瑶, 安雪姣. 沙雷氏菌(Serratia sp.) AXJ-M响应高温胁迫的生理适应性[J]. 微生物学通报, 2024.

https://doi.org/10.13344/j.microbiol.china.230940

WANG LW, WANG YL, HUI SY, FENG QQ, CHENG MLH, FENG R, LUO WY, LI YT, CHEN YY, AN XJ. Physiological adaptation of Serratia sp. AXJ-M to high temperature stress[J]. Microbiology China, 2024.

https://doi.org/10.13344/j.microbiol.china.230940

[42]

AN XJ, LI NJ, ZHANG SL, HAN YY, ZHANG QH. Integration of proteome and metabolome profiling to reveal heat stress response and tolerance mechanisms of Serratia sp. AXJ-M for the bioremediation of papermaking black liquor[J]. Journal of Hazardous Materials, 2023, 450:131092.

https://doi.org/10.1016/j.jhazmat.2023.131092

[43]

韩月颖, 张喜庆, 曲云鹏, 高云航. 一株低温木质素降解菌的筛选、产酶优化及酶学性质[J]. 微生物学通报, 2021, 48 (10):3700-3713.

HAN YY, ZHANG XQ, QV YP, GAO YH. Screening, enzyme-production optimization and enzymatic properties of a low-temperature lignin-degrading bacteria[J]. Microbiology China, 2021, 48 (10):3700-3713.

[44]

杨苹, 刘京, 周郑雄, 王坡, 谢世爽, 黄莺. 一株耐烟碱木质素降解菌的筛选及其对烟梗的降解效果[J]. 微生物学通报, 2024, 51 (6):2182-2192.

YANG P, LIU J, ZHOU ZX, WANG P, XIE SS, HUANG Y. A lignin-degrading strain with nicotine tolerance: screening and the degradation effect on tobacco stems[J]. Microbiology China, 2024, 51 (6):2182-2192.

[45]

庄培文, 张恺, 郑平, 凌铭煌, 张玉苍, 骆祝华. 深海真菌Chaetomium sp. CS1的木质素降解特征[J]. 菌物学报, 2023, 42 (12):2442-2453.

ZHUANG PW, ZHANG K, ZHENG P, LING MH, ZHANG YC, LUO ZH. Lignin-degradation characteristics of deep-sea fungus Chaetomium sp. CS1[J]. Mycosystema, 2023, 42 (12):2442-2453.

[46]

彭倩楠, 林璐. 木质素在海洋中的生物转化及其对海洋碳循环的影响[J]. 微生物学报, 2020, 60 (9):1959-1971.

https://doi.org/10.13343/j.cnki.wsxb.20200274

PENG QN, LIN L. Biotransformation of lignin in the ocean and its impact on marine carbon cycle[J]. Acta Microbiologica Sinica, 2020, 60 (9):1959-1971 (in Chinese).

https://doi.org/10.13343/j.cnki.wsxb.20200274

[47]

MARTIN-PASCUAL M, BATIANIS C, BRUINSMA L, ASIN-GARCIA E, GARCIA-MORALES L, WEUSTHUIS RA, van KRANENBURG R, MARTINS dos SANTOS VAP. A navigation guide of synthetic biology tools for Pseudomonas putida[J]. Biotechnology Advances, 2021, 49:107732.

https://doi.org/10.1016/j.biotechadv.2021.107732

[48]

YANG YR, SONG WY, HUR HG, KIM TY, GHATGE S. Thermoalkaliphilic laccase treatment for enhanced production of high-value benzaldehyde chemicals from lignin[J]. International Journal of Biological Macromolecules, 2019, 124:200-208.

https://doi.org/10.1016/j.ijbiomac.2018.11.144

[49]

KHAN SI, SAHINKAYA M, COLAK DN, ZADA NS, UZUNER U, BELDUZ AO, ÇANAKÇI S, KHAN AZ, KHAN S, BADSHAH M, SHAH AA. Production and characterization of novel thermostable CotA-laccase from Bacillus altitudinis SL7 and its application for lignin degradation[J]. Enzyme and Microbial Technology, 2024, 172:110329.

https://doi.org/10.1016/j.enzmictec.2023.110329

[50]

ZHU DC, LIANG N, ZHANG RX, AHMAD F, ZHANG WM, YANG B, WU J, GENG AL, GABRIEL M, SUN JZ. Insight into depolymerization mechanism of bacterial laccase for lignin[J]. ACS Sustainable Chemistry & Engineering, 2020, 8 (34):12920-12933.

[51]

CHAUHAN PS. Role of various bacterial enzymes in complete depolymerization of lignin: a review[J]. Biocatalysis and Agricultural Biotechnology, 2020, 23:101498.

https://doi.org/10.1016/j.bcab.2020.101498

[52]

VARDON DR, FRANDEN MA, JOHNSON CW, KARP EM, GUARNIERI MT, LINGER JG, SALM MJ, STRATHMANN TJ, BECKHAM GT. Correction: adipic acid production from lignin[J]. Energy & Environmental Science, 2022, 15 (8):3534-3535.

[53]

RATH S, PAUL M, BEHERA HK, THATOI H. Response surface methodology mediated optimization of lignin peroxidase from Bacillus mycoides isolated from Simlipal Biosphere Reserve, Odisha, India[J]. Journal of Genetic Engineering and Biotechnology, 2022, 20 (1):2.

https://doi.org/10.1186/s43141-021-00284-2

[54]

POLLEGIONI L, TONIN F, ROSINI E. Lignin-degrading enzymes[J]. The FEBS Journal, 2015, 282 (7):1190-1213.

https://doi.org/10.1111/febs.13224

[55]

QIN X, ZHANG J, ZHANG XY, YANG Y. Induction, purification and characterization of a novel manganese peroxidase from Irpex lacteus CD2 and its application in the decolorization of different types of dye[J]. PLoS One, 2014, 9 (11):e113282.

https://doi.org/10.1371/journal.pone.0113282

[56]

ZHANG XJ, ZHANG K, CHEN JC, YANG GH, JIA ZX. Enhancement of lignin removal from poplar prehydrolysis liquor by manganese peroxidase-induced polymerization[J]. Industrial Crops and Products, 2023, 206:117751.

https://doi.org/10.1016/j.indcrop.2023.117751

[57]

ZHANG ST, DONG ZJ, SHI J, YANG CR, FANG Y, CHEN G, CHEN H, TIAN CJ. Enzymatic hydrolysis of corn stover lignin by laccase, lignin peroxidase, and manganese peroxidase[J]. Bioresource Technology, 2022, 361:127699.

https://doi.org/10.1016/j.biortech.2022.127699

[58]

YU WN, LIU WN, HUANG HQ, ZHENG F, WANG XY, WU YY, LI KJ, XIE XM, JIN Y. Application of a novel alkali-tolerant thermostable DyP-type peroxidase from Saccharomonospora viridis DSM 43017 in biobleaching of eucalyptus kraft pulp[J]. PLoS One, 2014, 9 (10):e110319.

https://doi.org/10.1371/journal.pone.0110319

[59]

ELLEUCHE S, SCHÄFERS C, BLANK S, SCHRÖDER C, ANTRANIKIAN G. Exploration of extremophiles for high temperature biotechnological processes[J]. Current Opinion in Microbiology, 2015, 25:113-119.

https://doi.org/10.1016/j.mib.2015.05.011

[60]

VERMA JP, JAISWAL DK. Extremophilic enzymatic processing of lignocellulosic feedstocks to bioenergy: a book review[J]. Journal of Cleaner Production, 181:571-573.

https://doi.org/10.1016/j.jclepro.2018.02.026

[61]

AUSTIN S, KONTUR WS, ULBRICH A, OSHLAG JZ, ZHANG WP, HIGBEE A, ZHANG YP, COON JJ, HODGE DB, DONOHUE TJ, NOGUERA DR. Metabolism of multiple aromatic compounds in corn stover hydrolysate by Rhodopseudomonas palustris[J]. Environmental Science & Technology, 2015, 49 (14):8914-8922.

[62]

OKAI N, MASUDA T, TAKESHIMA Y, TANAKA K, YOSHIDA KI, MIYAMOTO M, OGINO C, KONDO A. Biotransformation of ferulic acid to protocatechuic acid by Corynebacterium glutamicum ATCC 21420 engineered to express vanillate O-demethylase[J]. AMB Express, 2017, 7 (1):130.

https://doi.org/10.1186/s13568-017-0427-9

[63]

FUCHS G, BOLL M, HEIDER J. Microbial degradation of aromatic compounds: from one strategy to four[J]. Nature Reviews Microbiology, 2011, 9:803-816.

https://doi.org/10.1038/nrmicro2652

[64]

KAMIMURA N, TAKAHASHI K, MORI K, ARAKI T, FUJITA M, HIGUCHI Y, MASAI EJ. Bacterial catabolism of lignin-derived aromatics: new findings in a recent decade: update on bacterial lignin catabolism[J]. Environmental Microbiology Reports, 2017, 9 (6):679-705.

https://doi.org/10.1111/1758-2229.12597

[65]

VARMAN AM, HE L, FOLLENFANT R, WU WH, WEMMER S, WROBEL SA, TANG YJ, SINGH S. Decoding how a soil bacterium extracts building blocks and metabolic energy from ligninolysis provides road map for lignin valorization[J]. Proceedings of the National Academy of Sciences of the United States of America, 2016, 113 (40):E5802-E5811.

[66]

AN XJ, CHENG Y, ZANG HL, LI CY. Biodegradation characteristics of lignin in pulping wastewater by the thermophilic Serratia sp. AXJ-M: performance, genetic background, metabolic pathway and toxicity assessment[J]. Environmental Pollution, 2023, 322:121230.

https://doi.org/10.1016/j.envpol.2023.121230

[67]

dos SANTOS MELO-NASCIMENTO AO, MOTA MOITINHO SANT ANNA B, GONÇALVES CC, SANTOS G, NORONHA E, PARACHIN N, de ABREU ROQUE MR, BRUCE T. Complete genome reveals genetic repertoire and potential metabolic strategies involved in lignin degradation by environmental ligninolytic Klebsiella variicola P1CD1[J]. PLoS One, 2020, 15 (12):e0243739.

https://doi.org/10.1371/journal.pone.0243739

[68]

HAN YJ, CHANG X, XIANG H, FANG Y, HAO LZ, GU Y, HAN XY, ZHAO B, ZHAO ZJ, ZHAO CZ, ZHONG RZ. Exploring biomimetic potential of ruminant digestion strategies for lignocellulosic biomass utilization: a comprehensive review[J]. Renewable and Sustainable Energy Reviews, 2023, 188:113887.

https://doi.org/10.1016/j.rser.2023.113887

[69]

GARG D, PATEL N, RAWAT A, ROSADO AS. Cutting edge tools in the field of soil microbiology[J]. Current Research in Microbial Sciences, 2024, 6:100226.

https://doi.org/10.1016/j.crmicr.2024.100226

[70]

DENMAN SE, MORGAVI DP, McSWEENEY CS. Review: the application of omics to rumen microbiota function[J]. Animal: an International Journal of Animal Bioscience, 2018, 12 (s2):s233-s245.

[71]

NEVES ALA, CHEN Y, LÊ CAO KA, MANDAL S, SHARPTON TJ, McALLISTER T, GUAN LL. Taxonomic and functional assessment using metatranscriptomics reveals the effect of Angus cattle on rumen microbial signatures[J]. Animal: an International Journal of Animal Bioscience, 2020, 14 (4):731-744.

https://doi.org/10.1017/S1751731119002453

[72]

HAN ML, ZHU Y, CREEK DJ, LIN YW, GUTU AD, HERTZOG P, PURCELL T, SHEN HH, MOSKOWITZ SM, VELKOV T, LI J. Comparative metabolomics and transcriptomics reveal multiple pathways associated with polymyxin killing in Pseudomonas aeruginosa[J]. mSystems, 2019, 4 (1):e00149-18.

[73]

ROSNOW JJ, ANDERSON LN, NAIR RN, BAKER ES, WRIGHT AT. Profiling microbial lignocellulose degradation and utilization by emergent omics technologies[J]. Critical Reviews in Biotechnology, 2017, 37 (5):626-640.

https://doi.org/10.1080/07388551.2016.1209158

[74]

LIEW SM, PUTHUCHEARY SD, RAJASEKARAM G, CHAI HC, CHUA KH. Proteomic profiling of clinical and environmental strains of Pseudomonas aeruginosa[J]. Molecular Biology Reports, 2021, 48 (3):2325-2333.

https://doi.org/10.1007/s11033-021-06262-8

[75]

AZAD RK, SHULAEV V. Metabolomics technology and bioinformatics for precision medicine[J]. Briefings in Bioinformatics, 2019, 20 (6):1957-1971.

https://doi.org/10.1093/bib/bbx170

[76]

VOHRA M, KOUR A, KALIA NP, KUMAR M, SHARMA S, JAGLAN S, KAMATH N, SHARMA S. A comprehensive review of genomics, transcriptomics, proteomics, and metabolomic insights into the differentiation of Pseudomonas aeruginosa from the planktonic to biofilm state: a multi-omics approach[J]. International Journal of Biological Macromolecules, 2024, 257 (Pt 1):128563.

[77]

HASSA J, MAUS I, OFF S, PÜHLER A, SCHERER P, KLOCKE M, SCHLÜTER A. Metagenome, metatranscriptome, and metaproteome approaches unraveled compositions and functional relationships of microbial communities residing in biogas plants[J]. Applied Microbiology and Biotechnology, 2018, 102 (12):5045-5063.

https://doi.org/10.1007/s00253-018-8976-7

[78]

TANG H, LI YQ, WANG MJ, LUO CB. Integrative microbial and transcriptomic analysis reveals the lignocellulosic biomass degrading mechanism by bamboo snout beetle[J]. Industrial Crops and Products, 2023, 203:117194.

https://doi.org/10.1016/j.indcrop.2023.117194

[79]

王一然, 康志超, 朱国鹏, 王洋, 其格其, 于洪文. 耐低温玉米秸秆降解菌群的优化及其效果[J]. 浙江农业学报, 2022, 34 (12):2720-2727.

WANG YR, KANG ZC, ZHU GP, WANG Y, QI GQ, YU HW. Optimization of low temperature resistant corn stalk degrading bacterial community and its effect[J]. Acta Agriculturae Zhejiangensis, 2022, 34 (12):2720-2727.

[80]

WANG M, LIU Y, JIANG XR, FANG JN, LYU QY, WANG XH, YAN ZY. Multi-omics reveal the structure and function of microbial community in co-digestion of corn straw and pig manure[J]. Journal of Cleaner Production, 2021, 322:129150.

https://doi.org/10.1016/j.jclepro.2021.129150

[81]

D'HAESELEER P, GLADDEN JM, ALLGAIER M, CHAIN PSG, TRINGE SG, MALFATTI SA, ALDRICH JT, NICORA CD, ROBINSON EW, PAŠA-TOLIĆ L, HUGENHOLTZ P, SIMMONS BA, SINGER SW. Proteogenomic analysis of a thermophilic bacterial consortium adapted to deconstruct switchgrass[J]. PLoS One, 2013, 8 (7):e68465.

https://doi.org/10.1371/journal.pone.0068465

[82]

MARTIEN JI, AMADOR-NOGUEZ D. Recent applications of metabolomics to advance microbial biofuel production[J]. Current Opinion in Biotechnology, 2017, 43:118-126.

https://doi.org/10.1016/j.copbio.2016.11.006

[83]

PAUL M, PANDEY NK, BANERJEE A, SHROTI GK, TOMER P, GAZARA RK, THATOI H, BHASKAR T, HAZRA S, GHOSH D. An insight into omics analysis and metabolic pathway engineering of lignin-degrading enzymes for enhanced lignin valorization[J]. Bioresource Technology, 2023, 379:129045.

https://doi.org/10.1016/j.biortech.2023.129045

[84]

SHU WS, HUANG LN. Microbial diversity in extreme environments[J]. Nature Reviews Microbiology, 2022, 20:219-235.

https://doi.org/10.1038/s41579-021-00648-y

[85]

RAFEEQ H, AFSHEEN N, RAFIQUE S, ARSHAD A, INTISAR M, HUSSAIN A, BILAL M, IQBAL HMN. Genetically engineered microorganisms for environmental remediation[J]. Chemosphere, 2023, 310:136751.

https://doi.org/10.1016/j.chemosphere.2022.136751

[86]

ISHAQ AR, ZHANG Z, HE PH, XIONG M, CHEN SW. Recent advances in keratinase production via protein engineering, breeding, and fermentation[J]. Advanced Agrochem, 2023: S2773237123001028.

[87]

邵明威, 孙思勉, 杨时茂, 陈国强. 基于极端微生物的生物制造[J]. 合成生物学, 2024.

https://doi.org/10.12211/2096-8280.2024-016

SHAO MW, SUN SM, YANG SM, CHEN GQ. Bioproduction based on extremophiles[J]. Synthetic Biology Journal, 2024.

https://doi.org/10.12211/2096-8280.2024-016

[88]

ZHAO YF, YANG J, WU YQ, HUANG BJ, XU LB, YANG JM, LIANG B, HAN L. Construction of bacterial laccase displayed on the microbial surface for ultrasensitive biosensing of phenolic pollutants with nanohybrids-enhanced performance[J]. Journal of Hazardous Materials, 2023, 452:131265.

https://doi.org/10.1016/j.jhazmat.2023.131265

2024年第64卷第11期

PDF下载

103

引用本文

BibTeX

文章信息

doi: 10.13343/j.cnki.wsxb.20240261

接收时间：2024-04-25
首发时间：2026-03-21
出版时间：2024-07-24

补充材料

相关文章

文章信息

作者

出版历史

收稿日期：2024-04-25
录用日期：2024-07-22

基金

Special Fund Project for High Tech Industrialization of Science and Technology Cooperation between Jilin Province and Chinese Academy of Sciences(2023SYHZ0008)

吉林省与中国科学院科技合作高技术产业化专项基金(2023SYHZ0008)

作者信息

中国科学院东北地理与农业生态研究所, 黑土地保护与利用全国重点实验室, 湿地生态与环境重点实验室, 吉林长春 130102

通讯作者:

参考文献

分享链接

https://castjournals.cast.org.cn/joweb/wswxb/CN/10.13343/j.cnki.wsxb.20240261

分享至

全文二维码

扫描看全文

引用本文

BibTeX

本文的引用情况

2种不同金属材料的力学参数

科 Family	属数 Number of genus	种数 Number of species	占总种数比例 Percentage of total species (%)	属 Genus	种数 Number of species	占总种数比例 Percentage of total species (%)
鹅膏菌科Amanitaceae	2	11	5.26	鹅膏菌属 Amanita	10	4.78
小菇科 Mycenaceae	2	12	5.74	丝盖伞属 Inocybe	5	2.39
多孔菌科 Polyporaceae	8	14	6.70	蜡蘑属 Laccaria	5	2.39
红菇科 Russulaceae	3	23	11.00	小皮伞属 Marasmius	6	2.87
				小菇属 Mycena	11	5.26
				光柄菇属 Pluteus	5	2.39
				红菇属 Russula	17	8.13
				栓菌属 Trametes	5	2.39

关闭全屏

BibTeX
EndNote
RefWorks
TxT

Species	Lignin (%)	Cinnamyl alcohol precursor (%)
		p-coumaryl alcohol	Coniferyl alcohol	Sinapyl alcohol
Hardwood	25−38	0−8	25−50	45−75
Softwood	20−30	< 5	> 95	0
Herbaceous plant	8−15	5−35	35−80	20−55

Species

Lignin (%)

Cinnamyl alcohol precursor (%)

p-coumaryl alcohol

Coniferyl alcohol

Sinapyl alcohol

Hardwood

25−38

0−8

25−50

45−75

Softwood

20−30

< 5

> 95

Herbaceous plant

8−15

5−35

35−80

20−55

Lignin-degrading strains	Growth characteristics	Source	References
Aneurinibacillus sp. LD3	Degradation of lignin up to 61.28% at 50 ℃	Soil samples covered with sawdust	[21]
Bacillus amyloliquefaciens SL-7	The strain can be composted for 13 days at temperatures between 55 ℃ and 62 ℃	Tobacco straw	[39]
Bacillus amyloliquefaciens HZ11	Degradation of lignin up to 46.7% at 50 ℃	Fungus chaff, straw and cow dung composted to produce seedling substrate	[40]
Serratia sp. AXJ-M	Strain was able to survive in heat stress environments ranging from 50−80 ℃, with 50 ℃ being the optimal temperature for growth	Soil	[41-42]
Stenotrophomonas maltophilia LS-1	The optimal conditions for enzyme production by the strain were pH 8.0 and temperature 15 ℃	Frozen soil	[43]
Arthrobacter sp. C2	The optimal conditions for enzyme production activity of this strain were initial pH 6.7 and temperature 14.9 ℃	Frozen soil	[22]
Enterobacter sp. YP-2023-9	The strain can tolerate at least 0.60% nicotine	Cow dung	[44]
Chaetomium sp. CS1	Promoting the release of nicotinic actives from tobacco stover	Bottomless sea sediment	[45]
Sutcliffiella sp. NC1	Alkaliphilic bacteria, which do not survive at pH 6.0, grow normally at pH 8.0−11.0	Soil	[24]
Bacillus ligniniphilus L1	The strain has an optimum growth temperature of 30 ℃, an optimum pH of 9.0 and can grow at 0−10% total salt concentration, with an optimum salt concentration of 2% (W/V)	Bottomless sea sediment	[26]
Pseudomonas putida KT2440	This strain is well adapted to salt osmotic stress, and at the same time has remarkable metabolic diversity and withstands difficult redox reactions and operational stresses	Soil	[46-47]

Lignin-degrading strains

Growth characteristics

Source

References

Aneurinibacillus sp. LD3

Degradation of lignin up to 61.28% at 50 ℃

Soil samples covered with sawdust

[21]

Bacillus amyloliquefaciens SL-7

The strain can be composted for 13 days at temperatures between 55 ℃ and 62 ℃

Tobacco straw

[39]

Bacillus amyloliquefaciens HZ11

Degradation of lignin up to 46.7% at 50 ℃

Fungus chaff, straw and cow dung composted to produce seedling substrate

[40]

Serratia sp. AXJ-M

Strain was able to survive in heat stress environments ranging from 50−80 ℃, with 50 ℃ being the optimal temperature for growth

Soil

[41-42]

Stenotrophomonas maltophilia LS-1

The optimal conditions for enzyme production by the strain were pH 8.0 and temperature 15 ℃

Frozen soil

[43]

Arthrobacter sp. C2

The optimal conditions for enzyme production activity of this strain were initial pH 6.7 and temperature 14.9 ℃

Frozen soil

[22]

Enterobacter sp. YP-2023-9

The strain can tolerate at least 0.60% nicotine

Cow dung

[44]

Chaetomium sp. CS1

Promoting the release of nicotinic actives from tobacco stover

Bottomless sea sediment

[45]

Sutcliffiella sp. NC1

Alkaliphilic bacteria, which do not survive at pH 6.0, grow normally at pH 8.0−11.0

Soil

[24]

Bacillus ligniniphilus L1

The strain has an optimum growth temperature of 30 ℃, an optimum pH of 9.0 and can grow at 0−10% total salt concentration, with an optimum salt concentration of 2% (W/V)

Bottomless sea sediment

[26]

Pseudomonas putida KT2440

This strain is well adapted to salt osmotic stress, and at the same time has remarkable metabolic diversity and withstands difficult redox reactions and operational stresses

Soil

[46-47]