Biocatalytic production of D-p-hydroxyphenylglycine by optimizing protein expression and cell wall engineering in Escherichia coli

doi:10.1007/s00253-019-10155-z

CORC > 上海应用物理研究所 > 中国科学院上海应用物理研究所 > 中科院上海应用物理研究所2011-2017年

	Biocatalytic production of D-p-hydroxyphenylglycine by optimizing protein expression and cell wall engineering in Escherichia coli
	Liu, Y ; Zhu, LF ; Qi, WP ; Yu, B
刊名	APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
	2019
卷号	103 期号:21-22 页码:8839-8851
关键词	AMINO ACID AMIDOHYDROLASE THERMOSTABLE D-HYDANTOINASE PENICILLIN-BINDING PROTEINS CRYSTAL-STRUCTURE BACILLUS-STEAROTHERMOPHILUS PSEUDOMONAS-PUTIDA OUTER-MEMBRANE WHOLE CELLS PEPTIDOGLYCAN ENZYME
ISSN号	0175-7598
DOI	10.1007/s00253-019-10155-z
文献子类	期刊论文
英文摘要	D-p-hydroxyphenylglycine (D-HPG) functions as an intermediate and has important value in antibiotic industries. The high pollution and costs from chemical processes make biotechnological route for D-HPG highly desirable. Here, a whole-cell transformation process by D-hydantoinase(Hase) and D-carbamoylase(Case) was developed to produce D-HPG from DL-hydroxyphenylhydantoin(DL-HPH) in Escherichia coli. The artificially designed ribosome binding site with strong intensity significantly facilitated the protein expression of limiting step enzyme Case. Next, the cell wall permeability was improved by disturbing the peptidoglycan structure by overproduction of D,D-carboxypeptidases without obviously affecting cell growth, to increase the bioavailability of low soluble hydantoin substrate. By fine-tuning regulation of expression level of D,D-carboxypeptidase DacB, the final production yield of D-HPG increased to 100% with 140 mM DL-HPH substrate under the optimized transformation conditions. This is the first example to enhance bio-productivity of chemicals by cell wall engineering and creates a new vision on biotransformation of sparingly soluble substrates. Additionally, the newly demonstrated 'hydroxyl occupancy' phenomenon when Case reacts with hydroxyl substrates provides a referential information for the enzyme engineering in future.
语种	英语
内容类型	期刊论文
源URL	[http://ir.sinap.ac.cn/handle/331007/31977]
专题	上海应用物理研究所_中科院上海应用物理研究所2011-2017年
作者单位	1.Forschungszentrum Julich, Inst Bio & Geosci, IBG 1 Biotechnol, Julich, Germany; 2.China Thailand Joint Lab Microbial Biotechnol, Beijing 100101, Peoples R China 3.Chinese Acad Sci, Inst Microbiol, CAS Key Lab Microbial Physiol & Metab Engn, State Key Lab Mycol, Beijing 100101, Peoples R China; 4.Univ Chinese Acad Sci, Beijing 100049, Peoples R China; 5.Chinese Acad Sci, Shanghai Inst Appl Phys, Shanghai 201800, Peoples R China;
推荐引用方式 GB/T 7714	Liu, Y,Zhu, LF,Qi, WP,et al. Biocatalytic production of D-p-hydroxyphenylglycine by optimizing protein expression and cell wall engineering in Escherichia coli[J]. APPLIED MICROBIOLOGY AND BIOTECHNOLOGY,2019,103(21-22):8839-8851.
APA	Liu, Y,Zhu, LF,Qi, WP,&Yu, B.(2019).Biocatalytic production of D-p-hydroxyphenylglycine by optimizing protein expression and cell wall engineering in Escherichia coli.APPLIED MICROBIOLOGY AND BIOTECHNOLOGY,103(21-22),8839-8851.
MLA	Liu, Y,et al."Biocatalytic production of D-p-hydroxyphenylglycine by optimizing protein expression and cell wall engineering in Escherichia coli".APPLIED MICROBIOLOGY AND BIOTECHNOLOGY 103.21-22(2019):8839-8851.