体外多酶分子机器产氢应用中的氢酶研究

李怡霏; 陈艾; 孙俊松; 张以恒

doi:10.12211/2096-8280.2024-052

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体外多酶分子机器产氢应用中的氢酶研究

特约评述 | 更新时间：2025-04-11

- 体外多酶分子机器产氢应用中的氢酶研究
- Studies on hydrogenases for hydrogen production using in vitro synthetic enzymatic biosystems
- 合成生物学 2024年5卷第6期页码：1461-1484
- 作者机构：
  
  1.中国科学院上海高等研究院低碳生物转化团队，上海 201210
  2.中国科学院大学，北京 100049
  3.中国科学院天津工业生物技术研究所低碳合成工程生物学（全国）重点实验室，天津 300308
  4.中国科学院天津工业生物技术研究所体外合成生物学中心，天津 300308
  5.合成生物学海河实验室，天津 300308
- 作者简介：
  
  [ "李怡霏（2000—），女，硕士研究生。研究方向为氢酶参与的体外多酶分子机器构建。 E-mail：liyf@sari.ac.cn" ]
  [ "孙俊松（1974—），男，博士，研究员。研究方向为氢酶表达、微生物代谢改造及生物合成。 E-mail：sunjs@sari.ac.cn" ]
  [ "张以恒（1971—），男，博士，研究员，中国科学院天津工业生物技术研究所低碳合成工程生物学（全国）重点实验室主任，曾任美国弗吉尼亚理工大学终身正教授。研究方向为体外合成生物学、新质生物制造、生物炼制和淀粉储能。 E-mail：zhang_xw@tib.cas.cn" ]
- 基金信息：
  
  国家重点研发计划“合成生物学”重点专项“糖水氢电系统——体外多酶高效产氢及氢电装置的基础及工程研究”(2022YFA0912000)
- DOI：10.12211/2096-8280.2024-052
  中图分类号： Q819
- 收稿：2024-07-09，
  
  修回：2024-09-25，
  
  纸质出版：2024-12-31
- 稿件说明：
移动端阅览
李怡霏，陈艾，孙俊松，张以恒. 体外多酶分子机器产氢应用中的氢酶研究［J］. 合成生物学， 2024， 5（6）： 1461-1484

LI Yifei， CHEN Ai， SUN Junsong， ZHANG Yi-Heng P. Job. Studies on hydrogenases for hydrogen production using in vitro synthetic enzymatic biosystems［J］. Synthetic Biology Journal， 2024， 5（6）： 1461-1484
李怡霏，陈艾，孙俊松，张以恒. 体外多酶分子机器产氢应用中的氢酶研究［J］. 合成生物学， 2024， 5（6）： 1461-1484 DOI： 10.12211/2096-8280.2024-052.

LI Yifei， CHEN Ai， SUN Junsong， ZHANG Yi-Heng P. Job. Studies on hydrogenases for hydrogen production using in vitro synthetic enzymatic biosystems［J］. Synthetic Biology Journal， 2024， 5（6）： 1461-1484 DOI： 10.12211/2096-8280.2024-052.

摘要

氢酶是生物制氢和氢能利用的最关键酶，它是一类广泛分布的对氧敏感的多亚基金属复合酶。体外多酶分子机器是体外生物转化技术中的高效酶生物催化系统，利用该分子机器生产氢气是一种新型高效的绿氢生产技术，它突破微生物产氢的Thauer极限，将葡萄糖产氢的转化率提高至接近化学理论值（1 mol葡萄糖裂解水生产12 mol氢气），代表着生物产氢的未来方向。氢酶的制备及催化性能是限制多酶分子机器产氢技术广泛应用的主要瓶颈；氧气不仅抑制氢酶的活性，也是氢酶转录翻译及翻译后加工的重要影响因素。体外多酶分子机器对氢酶的耐氧性能、热稳定性及高周转性能等参数提出高要求。本文结合氢酶在多酶分子机器制氢应用中的技术障碍，针对迫切的基础科学问题，分别从氢酶分类、结构功能、重组表达技术进展、（仿生）辅酶的适配等方面对其进行了相关的总结，并初步对氧的抑制机制、微生物重组表达氢酶以及产氢人工电子传递链的优化等难点问题的研究进行了跟踪，期待能够为氢酶在体外合成生物学的应用提供参考。

Abstract

Hydrogenases are the most important enzymes in biological hydrogen production and hydrogen energy utilization. They are widely distributed

oxygen-sensitive

multiunit complexed metal enzymes.

In vitro

synthetic enzymatic biosystems (ivSEB) is a type of

in vitro

biotransformation (ivBT) technology

which is an emerging biomanufacturing powerhouse that combines microbial fermentation with enzymatic biocatalysis

allowing for novel and efficient hydrogen production

also breaking the Thauer limit and achieving a yield of hydrogen close to the theoretical value of chemistry (1 mole of glucose to produce 12 moles of hydrogen in maximum). It represents the future direction of biological hydrogen production. However

the recombinant expression of hydrogenase is the main bottleneck limiting the wide application of ivSEB for hydrogen production technology. Hydrogenases are widely distributed in all life domains

but are oxygen-sensitive and mostly consist of metalloproteins with multi-subunits

bearing [Fe

]

only

[NiFe

]

or [FeFe

]

dinuclear core in their catalytic center. Oxygen not only inhibits the activity of hydrogenase

but also affects the transcription of the enzyme-encoding gene and post-translational process of the enzymes. As a result

the levels of recombinant hydrogenase are usually low and the enzymatic activitie

s are also incomparable to the native enzymes

often leading to high production costs due to the strict anaerobic purification procedures. In order to meet the requirements of industrial hydrogen production

hydrogenases must possess excellent catalytic properties

such as a high catalytic turnover number

great thermal stability

and the ability to tolerate trace amounts of oxygen. This review summarizes the studies on the structural and catalytic characterizations of hydrogenases

including their classification

oxygen resistance mechanisms

and progress in recombinant expression. Additionally

the evolution of natural electron transfer chains and the design of artificial routes

which can improve hydrogen production efficiency and reduce costs

are briefly discussed. The review also discussed the progress in the studies on the mechanisms of hydrogenases’ tolerance toward oxygen

the strategies for microbial expression of recombinant hydrogenases as well as the optimization of the artificial electron transfer chains adapted for the production of hydrogen using ivSEB

in expectations of promoting the applications of hydrogenases involved ivSEB

from renewable energy storage

anaerobic artificial respiration

to clean hydrogenation or dehydrogenation in biocatalysis.

关键词

Keywords

references

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