Metabolic engineering of Pseudomonas mendocina NK-01 for enhanced production of medium-chain-length polyhydroxyalkanoates with enriched content of the dominant monomer

被引：22

作者：

Zhao, Fengjie ^{[1
]}

He, Fanyang ^{[1
]}

Liu, Xiangsheng ^{[1
]}

Shi, Jie ^{[2
]}

Liang, Jingnan ^{[3
]}

Wang, Shufang ^{[2
]}

Yang, Chao ^{[1
]}

Liu, Ruihua ^{[2
]}

机构：

[1] Nankai Univ, Minist Educ, Key Lab Mol Microbiol & Technol, Tianjin 300071, Peoples R China

[2] Nankai Univ, State Key Lab Med Chem Biol, Tianjin 300071, Peoples R China

[3] Chinese Acad Sci, Inst Microbiol, Core Facil Equipment, Beijing 100101, Peoples R China

来源：

INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES | 2020年 / 154卷

关键词：

Pseudomonas mendocina; Metabolic engineering; mcl-PHA; FADA KNOCKOUT MUTANT; BETA-OXIDATION; CELL MORPHOLOGY; PHB PRODUCTION; PUTIDA; BIOSYNTHESIS; COPOLYMERS; SYNTHASES; GENOME; OPERON;

D O I：

10.1016/j.ijbiomac.2019.11.044

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

In this study, six genes involved in beta-oxidation pathway of P. mendocina NK-01 were deleted to construct mutant strains NKU-Delta beta 1 and NKU-Delta beta 5. Compared with the wild strain NKU, the mcl-PHA titers of NKU-Delta beta 5 were respectively increased by 5.58- and 4.85-fold for culturing with sodium octanoate and sodium decanoate. And the mcl-PHA titers of NKU-Delta beta 1 was increased by 10.02-fold for culturing with dodecanoic acid. The contents of dominant monomers 3-hydroxydecanoate (3HD) and 3-hydroxydodecanoate (3HDD) of the mcl-PHA synthesized by NKU-Delta beta 5 were obviously increased to 90.01 and 58.60 mol%, respectively. Further deletion of genes phaG and phaZ, the 3HD and 3HDD contents were further improved to 94.71 and 68.67 mol%, respectively. The highest molecular weight of mcl-PHA obtained in this study was 80.79 x 10(4) Da, which was higher than the previously reported mcl-PHA. With the increase of dominant monomer contents, the synthesized mcl-PHA showed better thermal properties, mechanical properties and crystallization properties. Interestingly, the cell size of NKU-Delta beta 5 was larger than that of NKU due to the accumulation of more PHA granules. This study indicated that a systematically metabolic engineering approach for P. mendocina NK-01 could significantly improve the mcl-PHA titer, dominant monomer contents and physical properties of mcl-PHA. (C) 2019 Published by Elsevier B.V.

引用

页码：1596 / 1605

页数：10

共 49 条

[21] Controlling cell volume for efficient PHB production by Halomonas [J].

Jiang, Xiao-Ran ;

Yao, Zhi-Hao ;

Chen, Guo-Qiang .

METABOLIC ENGINEERING, 2017, 44 :30-37

[22] Engineering the bacterial shapes for enhanced inclusion bodies accumulation [J].

Jiang, Xiao-Ran ;

Wang, Huan ;

Shen, Rui ;

Chen, Guo-Qiang .

METABOLIC ENGINEERING, 2015, 29 :227-237

[23] Biosynthesis and Properties of Medium-Chain-Length Polyhydroxyalkanoates with Enriched Content of the Dominant Monomer [J].

Jiang, Xuan ;

Sun, Zhiyong ;

Marchessault, Robert H. ;

Ramsay, Juliana A. ;

Ramsay, Bruce A. .

BIOMACROMOLECULES, 2012, 13 (09) :2926-2932

[24] Engineering Pseudomonas entomophila for synthesis of copolymers with defined fractions of 3-hydroxybutyrate and medium-chain-length 3-hydroxyalkanoates [J].

Li, Mengyi ;

Chen, Xiangbin ;

Che, Xuemei ;

Zhang, Haoqian ;

Wu, Lin-Ping ;

Du, Hetong ;

Chen, Guo-Qiang .

METABOLIC ENGINEERING, 2019, 52 :253-262

[25] The production of polyhydroxyalkanoates in recombinant Escherichia coli [J].

Li, Rui ;

Zhang, Hanxing ;

Qi, Qingsheng .

BIORESOURCE TECHNOLOGY, 2007, 98 (12) :2313-2320

[26] Microbial Synthesis of Functional Homo-, Random, and Block Polyhydroxyalkanoates by β-Oxidation Deleted Pseudomonas entomophila [J].

Li, Shijun ;

Cai, Longwei ;

Wu, Linping ;

Zeng, Guodong ;

Chen, Jinchun ;

Wu, Qiong ;

Chen, Guo-Qiang .

BIOMACROMOLECULES, 2014, 15 (06) :2310-2319

[27] Biosynthesis of poly(3-hydroxydecanoate) and 3-hydroxydodecanoate dominating polyhydroxyalkanoates by β-oxidation pathway inhibited Pseudomonas putida [J].

Liu, Qian ;

Luo, Ge ;

Zhou, Xin Rong ;

Chen, Guo-Qiang .

METABOLIC ENGINEERING, 2011, 13 (01) :11-17

[28] Production and characterization of medium-chain-length polyhydroxyalkanoate with high 3-hydroxytetradecanoate monomer content by fadB and fadA knockout mutant of Pseudomonas putida KT2442 [J].

Liu, Wenkuan ;

Chen, Guo-Qiang .

APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 2007, 76 (05) :1153-1159

[29] Production of two monomer structures containing medium-chain-length polyhydroxyalkanoates by β-oxidation-impaired mutant of Pseudomonas putida KT2442 [J].

Ma, Long ;

Zhang, Haifeng ;

Liu, Qian ;

Chen, Jiong ;

Zhang, Jing ;

Chen, Guo-Qiang .

BIORESOURCE TECHNOLOGY, 2009, 100 (20) :4891-4894

[30] Preparation and properties of a novel class of polyhydroxyalkanoate copolymers [J].

Noda, I ;

Green, PR ;

Satkowski, MM ;

Schechtman, LA .

BIOMACROMOLECULES, 2005, 6 (02) :580-586

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