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Article ; Online: CRISPR/Cpf1-FOKI-induced gene editing in

Wang, Xuyang / Li, Dong / Qin, Zhijie / Chen, Jian / Zhou, Jingwen

Synthetic and systems biotechnology

2024 Volume 9, Issue 2, Page(s) 369–379

Abstract: Gluconobacter ... ...

Abstract	Gluconobacter oxydans
Language	English
Publishing date	2024-03-04
Publishing country	China
Document type	Journal Article
ISSN	2405-805X
ISSN (online)	2405-805X
DOI	10.1016/j.synbio.2024.02.009
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Article ; Online: Combining Metabolic Engineering and Lipid Droplet Assembly to Achieve Campesterol Overproduction in

Qin, Zhijie / Zhang, Yunliang / Liu, Song / Zeng, Weizhu / Zhou, Jingwen / Xu, Sha

Journal of agricultural and food chemistry

2024 Volume 72, Issue 9, Page(s) 4814–4824

Abstract: Campesterol is a kind of important functional food additive. Therefore, stable and efficient campesterol biosynthesis is significant. Herein, we first knocked out the sterol 22-desaturase gene ... ...

Abstract	Campesterol is a kind of important functional food additive. Therefore, stable and efficient campesterol biosynthesis is significant. Herein, we first knocked out the sterol 22-desaturase gene in
MeSH term(s)	Saccharomyces cerevisiae/metabolism ; Metabolic Engineering ; Lipid Droplets/metabolism ; Saccharomyces cerevisiae Proteins/metabolism ; Sterols/metabolism ; Oxidoreductases/metabolism ; Cholesterol/analogs & derivatives ; Phytosterols
Chemical Substances	campesterol (5L5O665639) ; Saccharomyces cerevisiae Proteins ; Sterols ; Oxidoreductases (EC 1.-) ; Cholesterol (97C5T2UQ7J) ; Phytosterols
Language	English
Publishing date	2024-02-22
Publishing country	United States
Document type	Journal Article
ZDB-ID	241619-0
ISSN	1520-5118 ; 0021-8561
ISSN (online)	1520-5118
ISSN	0021-8561
DOI	10.1021/acs.jafc.3c09764
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Article: Dehydrogenases of acetic acid bacteria

Qin, Zhijie / Yu, Shiqin / Chen, Jian / Zhou, Jingwen

Biotechnology advances. 2022 Jan., Feb., v. 54

2022

Abstract: Acetic acid bacteria (AAB) are a group of bacteria that can oxidize many substrates such as alcohols and sugar alcohols and play important roles in industrial biotechnology. A majority of industrial processes that involve AAB are related to their ... ...

Abstract	Acetic acid bacteria (AAB) are a group of bacteria that can oxidize many substrates such as alcohols and sugar alcohols and play important roles in industrial biotechnology. A majority of industrial processes that involve AAB are related to their dehydrogenases, including PQQ/FAD-dependent membrane-bound dehydrogenases and NAD(P)⁺-dependent cytoplasmic dehydrogenases. These cofactor-dependent dehydrogenases must effectively regenerate their cofactors in order to function continuously. For PQQ, FAD and NAD(P)⁺ alike, regeneration is directly or indirectly related to the electron transport chain (ETC) of AAB, which plays an important role in energy generation for aerobic cell growth. Furthermore, in changeable natural habitats, ETC components of AAB can be regulated so that the bacteria survive in different environments. Herein, the progressive cascade in an application of AAB, including key dehydrogenases involved in the application, regeneration of dehydrogenase cofactors, ETC coupling with cofactor regeneration and ETC regulation, is systematically reviewed and discussed. As they have great application value, a deep understanding of the mechanisms through which AAB function will not only promote their utilization and development but also provide a reference for engineering of other industrial strains.
Keywords	acetic acid ; biotechnology ; cell growth ; electron transport chain ; energy ; oxidoreductases
Language	English
Dates of publication	2022-01
Publishing place	Elsevier Inc.
Document type	Article
ZDB-ID	47165-3
ISSN	0734-9750
ISSN	0734-9750
DOI	10.1016/j.biotechadv.2021.107863
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Article ; Online: Synergetic engineering of

Ping, Jurong / Wang, Lian / Qin, Zhijie / Zhou, Zhemin / Zhou, Jingwen

Synthetic and systems biotechnology

2023 Volume 8, Issue 4, Page(s) 724–731

Abstract: l-Tyrosine, an aromatic non-essential amino acid, is the raw material for many important chemical products, including levodopa, resveratrol, and hydroxytyrosol. It is widely used in the food, drug, and chemical industries. There are many studies on the ... ...

Abstract	l-Tyrosine, an aromatic non-essential amino acid, is the raw material for many important chemical products, including levodopa, resveratrol, and hydroxytyrosol. It is widely used in the food, drug, and chemical industries. There are many studies on the synthesis of l-tyrosine by microorganisms, however, the low titer of l-tyrosine limited the industrial large-scale production. In order to enhance l-tyrosine production in
Language	English
Publishing date	2023-11-07
Publishing country	China
Document type	Journal Article
ISSN	2405-805X
ISSN (online)	2405-805X
DOI	10.1016/j.synbio.2023.10.005
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Article ; Online: Enhancing Glycosylation of Flavonoids by Engineering the Uridine Diphosphate Glucose Supply in

Liu, Shike / Li, Dong / Qin, Zhijie / Zeng, Weizhu / Zhou, Jingwen

Journal of agricultural and food chemistry

2023 Volume 71, Issue 46, Page(s) 17842–17851

Abstract: Glycosylation can enhance the solubility and stability of flavonoids. The main limitation of the glycosylation process is low intracellular uridine diphosphate glucose (UDPG) availability. This study aimed to create a glycosylation platform strain ... ...

Abstract	Glycosylation can enhance the solubility and stability of flavonoids. The main limitation of the glycosylation process is low intracellular uridine diphosphate glucose (UDPG) availability. This study aimed to create a glycosylation platform strain in
MeSH term(s)	Glycosylation ; Uridine Diphosphate Glucose/metabolism ; Apigenin/metabolism ; Escherichia coli/genetics ; Escherichia coli/metabolism ; Luteolin/metabolism ; Flavonoids/metabolism
Chemical Substances	Uridine Diphosphate Glucose (V50K1D7P4Y) ; Apigenin (7V515PI7F6) ; Luteolin (KUX1ZNC9J2) ; Flavonoids
Language	English
Publishing date	2023-11-08
Publishing country	United States
Document type	Journal Article
ZDB-ID	241619-0
ISSN	1520-5118 ; 0021-8561
ISSN (online)	1520-5118
ISSN	0021-8561
DOI	10.1021/acs.jafc.3c05264
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Article ; Online: Production of Natural Pigments Using Microorganisms.

Qin, Zhijie / Wang, Xinglong / Gao, Song / Li, Dong / Zhou, Jingwen

Journal of agricultural and food chemistry

2023 Volume 71, Issue 24, Page(s) 9243–9254

Abstract: Pigments are involved in many aspects of human life, including food, cosmetics, and textiles. At present, the pigment market is mainly occupied by synthetic pigments. However, synthetic pigments have gradually presented safety and environmental problems. ...

Abstract	Pigments are involved in many aspects of human life, including food, cosmetics, and textiles. At present, the pigment market is mainly occupied by synthetic pigments. However, synthetic pigments have gradually presented safety and environmental problems. Therefore, humans have begun to focus on the use of natural pigments. In contrast to the extraction of pigments from plants and animals, the production of natural pigments by microbial fermentation is not affected by season and region. This review highlights recent advances in microbial synthesis of natural pigments, classifying them into various groups, including flavonoids, isoprenoids, porphyrins, N-heterocyclics, polyketides, and others. The biosynthetic pathways for each group are elucidated along with the latest progress made in enhancing production efficiency for both natural and non-natural microorganisms. Additionally, challenges associated with economically producing natural pigments using microorganisms are also discussed. This review provides a reference for researchers to replace synthetic pigments with natural pigments.
MeSH term(s)	Animals ; Humans ; Terpenes/metabolism ; Plants/metabolism ; Biosynthetic Pathways ; Flavonoids/metabolism
Chemical Substances	Terpenes ; Flavonoids
Language	English
Publishing date	2023-06-12
Publishing country	United States
Document type	Journal Article ; Review
ZDB-ID	241619-0
ISSN	1520-5118 ; 0021-8561
ISSN (online)	1520-5118
ISSN	0021-8561
DOI	10.1021/acs.jafc.3c02222
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Article ; Online: Engineering Gluconobacter cerinus CGMCC 1.110 for direct 2-keto-L-gulonic acid production

Qin, Zhijie / Chen, Yue / Yu, Shiqin / Chen, Jian / Zhou, Jingwen

Appl Microbiol Biotechnol. 2023 Jan., v. 107, no. 1 p.153-162

2023

Abstract: Gluconobacter is a potential strain for single-step production of 2-keto-L-gulonic acid (2-KLG), which is the direct precursor of vitamin C. Three dehydrogenases, namely, sorbitol dehydrogenase (SLDH), sorbose dehydrogenase (SDH), and sorbosone ... ...

Abstract	Gluconobacter is a potential strain for single-step production of 2-keto-L-gulonic acid (2-KLG), which is the direct precursor of vitamin C. Three dehydrogenases, namely, sorbitol dehydrogenase (SLDH), sorbose dehydrogenase (SDH), and sorbosone dehydrogenase (SNDH), are involved in the production of 2-KLG from D-sorbitol. In the present study, the potential SNDH/SDH gene cluster in the strain Gluconobacter cerinus CGMCC 1.110 was mined by genome analysis, and its function in transforming L-sorbose to 2-KLG was verified. Proteomic analysis showed that the expression level of SNDH/SDH had a great influence on the titer of 2-KLG, and fermentation results showed that SDH was the rate-limiting enzyme. A systematic metabolic engineering process, which was theoretically suitable for increasing the titer of many products involving membrane-bound dehydrogenase from Gluconobacter, was then performed to improve the 2-KLG titer in G. cerinus CGMCC 1.110 from undetectable to 51.9 g/L in a 5-L bioreactor after fermentation optimization. The strategies used in this study may provide a reference for mining other potential applications of Gluconobacter. KEY POINTS: • The potential SNDH/SDH gene cluster in G. cerinus CGMCC 1.110 was mined. • A systematic engineering process was performed to improve the titer of 2-KLG. • The 2-KLG titer was successfully increased from undetectable to 51.9 g/L.
Keywords	Gluconobacter cerinus ; L-iditol dehydrogenase ; ascorbic acid ; bioreactors ; fermentation ; multigene family ; proteomics ; sequence analysis ; sorbitol ; sorbose ; sorbose dehydrogenase
Language	English
Dates of publication	2023-01
Size	p. 153-162.
Publishing place	Springer Berlin Heidelberg
Document type	Article ; Online
ZDB-ID	392453-1
ISSN	1432-0614 ; 0171-1741 ; 0175-7598
ISSN (online)	1432-0614
ISSN	0171-1741 ; 0175-7598
DOI	10.1007/s00253-022-12310-5
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Article ; Online: Combined engineering of l-sorbose dehydrogenase and fermentation optimization to increase 2-keto-l-gulonic acid production in Escherichia coli

Li, Dong / Wang, Xinglong / Qin, Zhijie / Yu, Shiqin / Chen, Jian / Zhou, Jingwen

Bioresource Technology. 2023 Mar., v. 372 p.128672-

2023

Abstract: One-step fermentation to produce 2-keto-l-gulonic acid (2-KLG), the precursor of vitamin C, is a long-term goal. Improvement of the enzyme's activity through engineering could benefit 2-KLG production. This study aimed to conduct a semi-rational design ... ...

Abstract	One-step fermentation to produce 2-keto-l-gulonic acid (2-KLG), the precursor of vitamin C, is a long-term goal. Improvement of the enzyme's activity through engineering could benefit 2-KLG production. This study aimed to conduct a semi-rational design of l-sorbose dehydrogenase (SDH) through structure-directed, to screen mutants that could enhance the 2-KLG titer. First, the predicted structure of SDH was obtained using AlphaFold2. The key mutation sites in the substrate pocket were identified by Ala scanning. Subsequently, the mutant V336I/V368A was obtained by iterative saturation mutagenesis, which increased the yield of 2-KLG 1.9-fold. Finally, 5.03 g/L of 2-KLG was obtained by a two-stage temperature control fermentation method, and the conversion rate was 50%. Furthermore, experiments showed that knockdown of the l-sorbose-associated phosphotransferase system delays 2-KLG production. The results show that the production of 2-KLG was effectively increased through a combination of SDH engineering and fermentation optimization.
Keywords	Escherichia coli ; ascorbic acid ; fermentation ; mutagenesis ; mutants ; oxidoreductases ; sorbose ; temperature ; 2-Keto-l-gulonic acid ; l-Sorbose dehydrogenase ; l-Sorbose phosphotransferase system ; Semi-rational design ; Two-stage temperature control
Language	English
Dates of publication	2023-03
Publishing place	Elsevier Ltd
Document type	Article ; Online
ZDB-ID	1065195-0
ISSN	1873-2976 ; 0960-8524
ISSN (online)	1873-2976
ISSN	0960-8524
DOI	10.1016/j.biortech.2023.128672
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Article ; Online: Efficient production of 2-keto-L-gulonic acid from D-glucose in Gluconobacter oxydans ATCC9937 by mining key enzyme and transporter

Li, Guang / Li, Dong / Zeng, Weizhu / Qin, Zhijie / Chen, Jian / Zhou, Jingwen

Bioresource Technology. 2023 Sept., v. 384 p.129316-

2023

Abstract: Direct production of 2-keto-L-gulonic acid (2-KLG, the precursor of vitamin C) from D-glucose through 2,5-diketo-D-gluconic acid (2,5-DKG) is a promising alternative route. To explore the pathway of producing 2-KLG from D-glucose, Gluconobacter oxydans ... ...

Abstract	Direct production of 2-keto-L-gulonic acid (2-KLG, the precursor of vitamin C) from D-glucose through 2,5-diketo-D-gluconic acid (2,5-DKG) is a promising alternative route. To explore the pathway of producing 2-KLG from D-glucose, Gluconobacter oxydans ATCC9937 was selected as a chassis strain. It was found that the chassis strain naturally has the ability to synthesize 2-KLG from D-glucose, and a new 2,5-DKG reductase (DKGR) was found on its genome. Several major issues limiting production were identified, including the insufficient catalytic capacity of DKGR, poor transmembrane movement of 2,5-DKG and imbalanced D-glucose consumption flux inside and outside of the host strain cells. By identifying novel DKGR and 2,5-DKG transporter, the whole 2-KLG biosynthesis pathway was systematically enhanced by balancing intracellular and extracellular D-glucose metabolic flux. The engineered strain produced 30.5 g/L 2-KLG with a conversion ratio of 39.0%. The results pave the way for a more economical large-scale fermentation process for vitamin C.
Keywords	Gluconobacter oxydans ; ascorbic acid ; biosynthesis ; fermentation ; genome ; glucose ; oxidoreductases ; 2-keto-L-gulonic acid ; 2,5-diketo-D-glucose ; 2,5-DKG reductase ; 2,5-DKG transporter ; vitamin C
Language	English
Dates of publication	2023-09
Publishing place	Elsevier Ltd
Document type	Article ; Online
ZDB-ID	1065195-0
ISSN	1873-2976 ; 0960-8524
ISSN (online)	1873-2976
ISSN	0960-8524
DOI	10.1016/j.biortech.2023.129316
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Article ; Online: Coordinating caffeic acid and salvianic acid A pathways for efficient production of rosmarinic acid in Escherichia coli

Wang, Lian / Wang, Huijing / Chen, Jianbin / Qin, Zhijie / Yu, Shiqin / Zhou, Jingwen

International Metabolic Engineering Society Metabolic Engineering. 2023 Mar., v. 76 p.29-38

2023

Abstract: Rosmarinic acid is a natural hydroxycinnamic acid ester used widely in the food and pharmaceutical industries. Although many attempts have been made to screen rate-limiting enzymes and optimize modules through co-culture fermentation, the titer of ... ...

Abstract	Rosmarinic acid is a natural hydroxycinnamic acid ester used widely in the food and pharmaceutical industries. Although many attempts have been made to screen rate-limiting enzymes and optimize modules through co-culture fermentation, the titer of rosmarinic acid remains at the microgram level by microorganisms. A de novo biosynthetic pathway for rosmarinic acid was constructed based on caffeic acid synthesis modules in Escherichia coli. Knockout of competing pathways increased the titer of rosmarinic acid and reduced the synthesis of rosmarinic acid analogues. An L-amino acid deaminase was introduced to balance metabolic flux between the synthesis of caffeic acid and salvianic acid A. The ratio of FADH₂/FAD was maintained via the coordination of deaminase and HpaBC, which is responsible for caffeic acid synthesis. Knockout of menI, encoding an endogenous thioesterase, increased the stability of caffeoyl-CoA. The final strain produced 5780.6 mg/L rosmarinic acid in fed-batch fermentation, the highest yet reported for microbial production. The strategies applied in this study lay a foundation for the synthesis of other caffeic acid and rosmarinic acid derivatives.
Keywords	Escherichia coli ; amino acids ; batch fermentation ; biochemical pathways ; caffeic acid ; coculture ; rosmarinic acid ; L-amino acid Deaminase ; Cofactor ; Thioesterase
Language	English
Dates of publication	2023-03
Size	p. 29-38.
Publishing place	Elsevier Inc.
Document type	Article ; Online
ZDB-ID	1470383-x
ISSN	1096-7184 ; 1096-7176
ISSN (online)	1096-7184
ISSN	1096-7176
DOI	10.1016/j.ymben.2023.01.002
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