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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
2019
Source PublicationAPPLIED MICROBIOLOGY AND BIOTECHNOLOGY
ISSN0175-7598
Volume103Issue:21-22Pages:8839-8851
Subtype期刊论文
AbstractD-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.
KeywordAMINO ACID AMIDOHYDROLASE THERMOSTABLE D-HYDANTOINASE PENICILLIN-BINDING PROTEINS CRYSTAL-STRUCTURE BACILLUS-STEAROTHERMOPHILUS PSEUDOMONAS-PUTIDA OUTER-MEMBRANE WHOLE CELLS PEPTIDOGLYCAN ENZYME
DOI10.1007/s00253-019-10155-z
Indexed BySCI
Language英语
Citation statistics
Document Type期刊论文
Identifierhttp://ir.sinap.ac.cn/handle/331007/31977
Collection中科院上海应用物理研究所2011-2020年
Affiliation1.Chinese Acad Sci, Inst Microbiol, CAS Key Lab Microbial Physiol & Metab Engn, State Key Lab Mycol, Beijing 100101, Peoples R China;
2.Univ Chinese Acad Sci, Beijing 100049, Peoples R China;
3.Forschungszentrum Julich, Inst Bio & Geosci, IBG 1 Biotechnol, Julich, Germany;
4.Chinese Acad Sci, Shanghai Inst Appl Phys, Shanghai 201800, Peoples R China;
5.China Thailand Joint Lab Microbial Biotechnol, Beijing 100101, Peoples R China
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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.
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