A combined strategy of metabolic pathway regulation and two-step bioprocess for improved 4-androstene-3,17-dione production with an engineered Mycobacterium neoaurum

被引：10

作者：

Chang, Hongxing ^{[1
]}

Zhang, Hongtao ^{[1
]}

Zhu, Lin ^{[1
]}

Zhang, Wei ^{[5
,6
]}

You, Shengping ^{[1
,4
]}

Qi, Wei ^{[1
,2
,3
,4
]}

Qian, Jianwu ^{[6
]}

Su, Rongxin ^{[1
,2
,3
,4
]}

He, Zhimin ^{[1
,2
]}

机构：

[1] Tianjin Univ, Chem Engn Res Ctr, Sch Chem Engn & Technol, Tianjin 300072, Peoples R China

[2] Tianjin Univ, State Key Lab Chem Engn, Tianjin 300072, Peoples R China

[3] Collaborat Innovat Ctr Chem Sci & Engn Tianjin, Tianjin 300072, Peoples R China

[4] Tianjin Univ, Tianjin Key Lab Membrane Sci & Desalinat Technol, Tianjin 300072, Peoples R China

[5] Ever Sky Biosci Tianjin Co Ltd, Tianjin, Peoples R China

[6] Biosyn Hlth Pharma Co Ltd, Tianjin, Peoples R China

来源：

BIOCHEMICAL ENGINEERING JOURNAL | 2020年 / 164卷

基金：

中国博士后科学基金;

关键词：

Mycobacterium neoaurum; Metabolic pathway regulation; Two-step bioprocess; 4-androstene-3; 17-dione; Phytosterols; RESTING CELLS; BIOTRANSFORMATION; PHYTOSTEROL; ANDROSTENEDIONE; DEGRADATION; CONVERSION; STEROLS; STEP;

D O I：

10.1016/j.bej.2020.107789

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

This study aims to improve the efficiency of 4-androstene-317-dione (AD) production from high-concentrated phytosterols through a combined strategy of metabolic pathway regulation and two-step bioprocess. Firstly, we overexpressed cholesterol oxidase and steroid C27 monooxygenase to strengthen the metabolic flux to AD and then enhanced the expression of 17 beta-hydroxystemid dehydrogenase to reduce the by-product 20-hydroxy-methylpregna-4-ene-3-one. As a result, the AD production was enhanced from 2.4 to 4.5 g/L with an increase of 88.6 %. Afterward, a two-step bioprocess, cell culture at 30 degrees C and bioconversion with resting cells at 37 degrees C, was adopted to avoid the cell growth inhibition and decrease the steroid nucleus degradation. Finally, 25.8 g/L AD was produced from 50 g/L phytosterols, which is significantly higher than that from the fermentation bioprocess and is the highest reported AD production. Therefore, this work hopes to pave the way for the commercial production of AD through Mycobacterium transformation.

引用

页数：8

共 34 条

[1] Conversion of soybean sterols into 3,17-diketosteroids using actinobacteria Mycobacterium neoaurum, Pimelobacter simplex, and Rhodococcus erythropolis [J].

Andryushina, V. A. ;

Rodina, N. V. ;

Stytsenko, T. S. ;

Luu Duc Huy ;

Druzhinina, A. V. ;

Yaderetz, V. V. ;

Voishvillo, N. E. .

APPLIED BIOCHEMISTRY AND MICROBIOLOGY, 2011, 47 (03) :270-273

[2] Production of tropane alkaloids by biotransformation using recombinant Escherichia coli whole cells [J].

Cardillo, Alejandra B. ;

Perassolo, Maria ;

Sartuqui, Mariela ;

Rodriguez Talou, Julian ;

Giulietti, Ana M. .

BIOCHEMICAL ENGINEERING JOURNAL, 2017, 125 :180-189

[3] Whole cell biocatalysts: essential workers from Nature to the industry [J].

de Carvalho, Carla C. C. R. .

MICROBIAL BIOTECHNOLOGY, 2017, 10 (02) :250-263

[4]

Donova M.V., 1996, MODIFIED CDS MEDIATE

[5] Microbial steroid transformations: current state and prospects [J].

Donova, Marina V. ;

Egorova, Olga V. .

APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 2012, 94 (06) :1423-1447

[6] Production of androstenedione using mutants of Mycobacterium sp [J].

Egorova, OV ;

Gulevskaya, SA ;

Puntus, IF ;

Filonov, AE ;

Donova, MV .

JOURNAL OF CHEMICAL TECHNOLOGY AND BIOTECHNOLOGY, 2002, 77 (02) :141-147

[7] Phytosterols: Applications and recovery methods [J].

Fernandes, P. ;

Cabral, J. M. S. .

BIORESOURCE TECHNOLOGY, 2007, 98 (12) :2335-2350

[8] New Insights on Steroid Biotechnology [J].

Fernandez-Cabezon, Lorena ;

Galan, Beatriz ;

Garcia, Jose L. .

FRONTIERS IN MICROBIOLOGY, 2018, 9

[9] Enhanced Steroid Metabolites Production by Resting Cell Phytosterol Bioconversion [J].

Gao, X. -Q. ;

Feng, J. -X. ;

Wang, X. -D. ;

Hua, Q. ;

Wei, D. -Z. .

CHEMICAL AND BIOCHEMICAL ENGINEERING QUARTERLY, 2015, 29 (04) :567-573

[10] Investigation of factors affecting biotransformation of phytosterols to 9-hydroxyandrost-4-ene-3,-17-dione based on the HP-β-CD-resting cells reaction system [J].

Gao, Xingqiang ;

Feng, Jianxun ;

Hua, Qiang ;

Wei, Dongzhi ;

Wang, Xuedong .

BIOCATALYSIS AND BIOTRANSFORMATION, 2014, 32 (5-6) :343-347

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