合成生物学助力废弃塑料资源生物解聚与升级再造

doi:10.12211/2096-8280.2020-087

合成生物学 ›› 2021, Vol. 2 ›› Issue (2): 161-180.DOI: 10.12211/2096-8280.2020-087

合成生物学助力废弃塑料资源生物解聚与升级再造

钱秀娟¹, 刘嘉唯¹, 薛瑞¹, 刘豪杰¹, 闻小红¹, 杨璐¹, 徐安明¹, 许斌¹, 信丰学¹^,², 周杰¹^,², 董维亮¹^,², 姜岷¹^,²

^1.南京工业大学生物与制药工程学院，江苏南京 211816
^2.南京工业大学材料化学工程国家重点实验室，江苏南京 211816

收稿日期:2020-12-04 修回日期:2021-02-11 出版日期:2021-04-29 发布日期:2021-04-30
通讯作者: 董维亮,姜岷
作者简介:钱秀娟（1992—），女，博士，博士后，研究方向为代谢工程及合成生物学。E-mail：xiujuanqian@njtech.edu.cn
董维亮（1988—），男，博士，教授，研究方向为环境污染物的生物降解与转化利用。E-mail：dwl@njtech.edu.cn
姜岷（1972—），男，博士，教授，研究方向为废弃碳资源利用人工多细胞体系设计与构建。E-mail：jiangmin@njtech.edu.cn
基金资助:
国家自然科学基金国际（地区）合作与交流项目(31961133017);国家重点研发计划“合成生物学”重点专项(2019YFA0905500);国家自然科学基金(21978129);江苏省农业自主创新计划(CX(19)3104);江苏省研究生科研与实践创新计划(KYCX20_1100)

Synthetic biology boosts biological depolymerization and upgrading of waste plastics

Xiujuan QIAN¹, Jiawei LIU¹, Rui XUE¹, Haojie LIU¹, Xiaohong WEN¹, Lu YANG¹, Anming XU¹, Bin XU¹, Fengxue XIN¹^,², Jie ZHOU¹^,², Weiliang DONG¹^,², Min JIANG¹^,²

^1.College of Biotechnology and Pharmaceutical Engineering，Nanjing Tech University，Nanjing 211816，Jiangsu，China
^2.State Key Laboratory of Materials-Oriented Chemical Engineering，Nanjing Tech University，Nanjing 211816，Jiangsu，China

Received:2020-12-04 Revised:2021-02-11 Online:2021-04-29 Published:2021-04-30
Contact: Weiliang DONG, Min JIANG

摘要/Abstract

摘要：

石油基合成塑料因高分子量、高疏水性及高化学键能等特性难以被生物降解，在环境中不断累积，由此导致的“白色污染”已经成为一个全球性环境问题。填埋和焚烧是目前塑料垃圾处置最简单、常用的方法，但随之带来的是更为严重的环境二次污染问题。为解决这一问题，开发绿色高效的废塑料资源回收利用技术，从源头解决塑料污染，成为发展塑料循环经济的关键。利用微生物/酶将塑料降解为寡聚体或单体，或进一步转化为高值化学品，因反应条件温和、不产生二次污染等优点将成为废塑料污染治理与资源化的新途径。本文详细介绍了废塑料生物解聚与转化方面的最新研究进展，包括塑料降解微生物和酶的挖掘、混菌/多酶体系的设计与构建、塑料解聚机制，以及塑料解聚物到化学品、能源、材料等高附加值产品的转化。然而，废塑料生物降解过程中仍存在降解元件匮乏、降解效率低、降解物难以利用等技术瓶颈。随着合成生物学的快速发展，利用高通量筛选、进化代谢、生物信息学等先进的生物技术，解析降解关键酶的催化机制、定向设计与改造降解酶、研究混菌体系中菌株间互利共生关系与适配机制、设计并构建不同塑料降解物的代谢通路成为废塑料生物降解研究的重点方向。通过建立废塑料生物降解与高值化利用平台，可为巨量的废塑料资源循环利用提供新的理论基础和关键技术，为我国塑料循环经济发展提供经济、环保、可行的技术支撑。

关键词: 废塑料, 生物解聚, 生物转化, 混菌/多酶, 升级再造

Abstract:

Characterics of high molecular weight, high hydrophobicity and high chemical bond energy make petroleum-based synthetic plastics resist to abiotic and microbial degradation. "White pollution" caused by accumulated waste plastics in the environment has become a global challenge. Currently, landfill and incineration are the simplest and most commonly used methods for eliminating plastic wastes, given that only 20% of plastic wastes is recycled, but landfill and incineration cause more serious secondary hazards, such as pollution to groundwater, soil, air and ocean. Therefore, developing a green and efficient technology for recycling and reutilization plastic wastes is the key for solving plastic pollution, to boost a plastic recycling economy.Applications of microorganisms/enzymes to degrade plastics into oligomers or monomers, which can be further transformed into high-value added chemicals, have provided a new approach for such a purpose due to their mild and environmentally friendly proceses. This article comprehensively reviews the development of biodepolymerization and biotransformation for waste plastics, including mining plastic degrading microorganisms and enzymes, designing and constructing microbial consortia/enzyme cocktails, analyzing of plastics depolymerization mechanism, and transforming plastics degradants into high value-added products, such as chemicals, energy products, and materials. However, the lack of degradation enzyme components, low degradation efficiency and difficulty for utilizing the degradants limit the development of waste plastics biogradation. With advances in synthetic biology, emerging technologies, such as high-throughput screening, evolutionary metabolism, and bioinformatics to analyze the catalytic mechanism of key degradation enzymes, orientedly designing and modification of degradation enzymes, study of the mutualism relationship and mechanism in the microbial consortia, constructing metabolic pathways for different plastic degradants, will open windows for waste plastics biodegradation, providing environmentally friendly， economically competitive and technically feasible technologies to develop circular economy for the re-utilization of waste plastics in China.

Key words: waste plastics, biological depolymerization, biotransformation, microbial consortia/enzyme cocktails, re-utilization

中图分类号:

Q812

钱秀娟, 刘嘉唯, 薛瑞, 刘豪杰, 闻小红, 杨璐, 徐安明, 许斌, 信丰学, 周杰, 董维亮, 姜岷. 合成生物学助力废弃塑料资源生物解聚与升级再造[J]. 合成生物学, 2021, 2(2): 161-180.

Xiujuan QIAN, Jiawei LIU, Rui XUE, Haojie LIU, Xiaohong WEN, Lu YANG, Anming XU, Bin XU, Fengxue XIN, Jie ZHOU, Weiliang DONG, Min JIANG. Synthetic biology boosts biological depolymerization and upgrading of waste plastics[J]. Synthetic Biology Journal, 2021, 2(2): 161-180.

图/表 9

图1 2019年中国废塑料处理情况[4]（According to 2019—2020 Development Report of China's Plastic Recycling Industry, in 2019, 134 million tons of primary plastic raw materials have been supplied in China, and the output of plastic products was 96 million tons. The generation of waste plastics was 63 million tons, of which 20 million tons went to landfill; 19 million tons was incinerated, 4 million tons was discarded, accounting for 30%, 32%, 31% and 7% of the total waste plastics, respectively. Only 19 million tons was recycled, accounting for 30% of the total plastics）

Fig. 1 Fates for China's waste plastics in 2019[4]

表1 水解型塑料降解微生物研究进展

Tab. 1 Microorganisms responsible for depolymerizing plastics through hydrolysis

塑料分类	降解菌	降解温度/℃	降解效果	文献
PET	Fusarium solani	30	PET纤维表面改性	[11-12]
	Humicola insolens	30	PET纤维表面改性	[13]
	Thermobifida fusca	30	PET纤维表面改性	[14-17]
	Saccharomonospora viridis	30	PET纤维表面改性	[18]
	Ideonella sakaiensis 201-F6	30	6周内能完全降解低结晶度PET薄膜	[19]
PU	Aspergillus flavus (ITCC 6051)	28±2	30 d降解60.6%聚酯型PU薄膜	[20]
	Aspergillus tubingensis	37	14 d降解聚酯型PU薄膜成碎片	[21]
	Aspergillus sp. strain S45	37	28 d降解20%聚酯型PU薄膜	[22]
	Cladosporium pseudocladosporioides, Cladosporium tenuissimum, Cladosporium asperulatum, Cladosporium montecillanum, Aspergillus fumigatus, Penicillium chrysogenum	25~30	21 d质降解10%～65%聚酯型PU薄膜	[23]

表2 水解型塑料解聚酶挖掘

Tab. 2 Depolymerases responsible for plastics depolymerization through hydrolysis

塑料分类	降解底物	解聚酶来源	解聚酶	降解温度/℃	降解能力	文献
PET	饮料瓶	Thermobifida fusca DSM43793	TfH	55	21 d质量损失50%	[27]
	低结晶度（7%）PET薄膜	Humicola insolens	HiC	70	96 h重量损失97%	[28]
	PET薄膜	plant compost	LCC	70	24 h重量损失25%	[29]
	低结晶度（1.9%）PET薄膜	Ideonella sakaiensis 201-F6	PETase	30	—	[29]
	低结晶度PET薄膜	T. fusca KW3	TfCut2	65～80	48 h重量损失12%	[30-31]
PU	Impranil DLN	Comamonas acidovorans TB-35	PudA	45	—	[32-33]
	Impranil DLN	Pseudomonas fluorescens	PulA	48	—	[34]
	Impranil DLN	Pseudomonas chlororaphis	PueA/PueB	65/60	—	[35-36]
	固体聚酯型PU	T. fusca KW3	TfCut2	70	100 h重量损失1.9%	[37]
	固体聚酯型PU	plant compost	LCC	70	100 h重量损失3.2%	[37]

表3 非水解型塑料降解微生物分离研究进展

Tab. 3 Microorganisms responsible for plastics depolymerization through non-hydrolysis pathways

塑料分类	降解底物	降解菌	降解温度/℃	降解效果	文献
PE	改性PE膜	Aspergillus niger M6	28	30 d质量损失20%	[44]
	高密度PE	Arthrobacter sp. GMB5	30	30 d质量损失12%	[45]
	高密度PE	Pseudomonas sp. GMB7	30	30 d质量损失15%	[45]
	低密度PE薄膜	Enterobacter asburiae YT1	30	60 d质量损失6.1%±0.3%	[46]
	低密度PE薄膜	Bacillus sp. YP1	30	60 d质量损失10.7%±0.2%	[46]
PS	PS泡沫	Mealworms (the larvae of Tenebrio molitor Linnaeus)	30	30 d质量损失31.0%±1.7%	[47]
	PS薄膜	Exiguobacterium sp. YT2	30	60 d质量损失7.4%±0.4%	[47]
	PS薄膜	Penicillium variabile	24	16周内能矿化完	[48]

表4 非水解型塑料解聚酶元件挖掘研究进展

Tab. 4 Depolymerases responsible for plastics depolymerization through non-hydrolysis pathways

塑料分类	降解底物	解聚酶来源	解聚酶	降解温度/℃	降解能力	文献
PE	氧化后的低密度PE	Phanerochaete chrysosporium MTCC-787	LiP/MnP	37	15 d内降解70%	[60]
PE	低分子量 PE粉末	Pseudomonas sp. E4	alkB	37	80 d内降解20%	[57]
	低分子量 PE粉末	Pseudomonas aeruginosa E7	AH系统	37	80 d内降解30%	[58]
PS	PS	Azotobacter beijerinckii HM121	非血红素氢醌过氧化物酶	30	5 min内水解PS转化为水溶性产物	[59]

图2 有机酸类塑料单体（己二酸和6-羟基己酸等）的生物降解途径（途径涉及的关键酶：DcaIJ—琥珀酰CoA转移酶；DcaA—酰基CoA脱氢酶；DcaE—烯酰CoA水合酶；DcaH—3-羟基己二酰CoA脱氢酶；DcaF—酰基CoA硫解酶；ChnD—6-羟基己酸脱氢酶；ChnE—6-氧己酸脱氢酶）

Fig. 2 Biological degradation pathway for plastics from organic acid based monomers (adipic acid, 6-hydroxyhexanoic acid, etc.)（Key enzymes in metabolic pathway: DcaIJ—succinyl-CoA transferase; DcaA—acyl-CoA dehydrogenase; DcaE—enoyl-CoA hydratase; DcaH—3-hyroxyacyl-CoA dehydrogenase; DcaF—acyl-CoA thiolase； ChnD—6-hydroxyhexanoate dehydrogenase; ChnE—6-oxohexanoate dehydrogenase)

图3 有机醇类塑料单体（乙二醇和1,4-丁二醇等）的生物降解途径（途径涉及的关键酶：PedE，PedH—醌依赖性醇脱氢酶； PP_0545和PedI—醛脱氢酶；GlcDEF—乙醇酸氧化酶；Gcl—乙醛酸羧化酶；Hyi—羟基丙酮酸异构酶；GlxR—酒石酸酯半醛还原酶；AceA—异柠檬酸裂解酶；GlcB—苹果酸合成酶；PduP—丙醛脱氢酶）

Fig. 3 Biological degradation pathway for plastic from organic alcohol based monomers (ethylene glycol, 1,4-butanediol， etc.)（Key enzymes in metabolic pathway:PedE, PedH—quinoprotein alcohol dehydrogenase; PP_0545 and PedI—aldehyde dehydrogenase; GlcDEF—glycolate oxidase; Gcl—glyoxylate carboligase; Hyi—hydroxypyruvate isomerase; GlxR—artronate semialdehyde reductase; AceA—isocitrate lyase; GlcB—malate synthase; PduP—propionaldehyde dehydrogenase）

图4 芳香类塑料单体（对苯二甲酸和2,4-二氨基甲苯等）的生物降解途径（途径涉及的关键酶：TphAabc—对苯二甲酸1,2-双加氧酶；TphB—1,2-二羟基-3,5-环己二烯-1,4-二羧酸酯脱氢酶）

Fig. 4 Biological degradation pathway for plastics from aromatic monomers (terephthalic acid, 2,4-diaminotoluene， etc.)（Key enzymes in metabolic pathway: TphAabc—TPA 1,2-dioxygenase; TphB—1,2-dihydroxy-3,5-cyclohexadiene-1,4-dicarboxylate dehydrogenase）

图5 脂肪烃类塑料单体的生物降解途径（途径涉及的关键酶：P450—单加氧酶P450；SDO—苯乙烯双加氧酶；SMO—苯乙烯单加氧酶；CGDH—顺式乙二醇脱氢酶；SOI—氧化苯乙烯异构酶；PAALDH—苯乙醛脱氢酶）

Fig. 5 Biological degradation pathway for plastics from aliphatic hydrocarbon monomers（Key enzymes in metabolic pathway: P450—monooxygenase P450; SDO—styrene dioxygenase; SMO—styrene monooxygenase; CGDH—cis-ethylene glycol dehydrogenase; SOI—styrene oxide isomerase; PAALDH—phenylacetaldehyde dehydrogenase）

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合成生物学助力废弃塑料资源生物解聚与升级再造

Synthetic biology boosts biological depolymerization and upgrading of waste plastics

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