scholarly journals Photosynthetic Conversion of CO2 Into Pinene Using Engineered Synechococcus sp. PCC 7002

2021 ◽  
Vol 9 ◽  
Author(s):  
Ruigang Yang ◽  
Lingyun Zhu ◽  
Tao Li ◽  
Lv-yun Zhu ◽  
Zi Ye ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Metabolic Engineering ◽  
Carbon Chain ◽  
Gene Encoding ◽  
Photoautotrophic Growth ◽  
Abies Grandis ◽  
Growth Performances ◽  
Synechococcus Sp ◽  
Suitable Potential ◽  
Geranyl Pyrophosphate

Metabolic engineering of cyanobacteria has received much attention as a sustainable strategy to convert CO2 to various longer carbon chain fuels. Pinene has become increasingly attractive since pinene dimers contain high volumetric energy and have been proposed to act as potential aircraft fuels. However, cyanobacteria cannot directly convert geranyl pyrophosphate into pinene due to the lack of endogenous pinene synthase. Herein, we integrated the gene encoding Abies grandis pinene synthase into the model cyanobacterium Synechococcus sp. PCC 7002 through homologous recombination. The genetically modified cyanobacteria achieved a pinene titer of 1.525 ± 0.l45 mg L−1 in the lab-scale tube photobioreactor with CO2 aeration. Specifically, the results showed a mixture of α- and β-pinene (∼33:67 ratio). The ratio of β-pinene in the product was significantly increased compared with that previously reported in the engineered Escherichia coli. Furthermore, we investigated the photoautotrophic growth performances of Synechococcus overlaid with different concentrations of dodecane. The work demonstrates that the engineered Synechococcus is a suitable potential platform for β-pinene production.

scholarly journals Combining protein and metabolic engineering strategies for biosynthesis of melatonin in Escherichia coli

2021 ◽  
Vol 20 (1) ◽  
Author(s):  
Yanfeng Zhang ◽  
Yongzhi He ◽  
Nan Zhang ◽  
JiaJia Gan ◽  
Shan Zhang ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Metabolic Engineering ◽  
Xanthomonas Campestris ◽  
Crop Improvement ◽  
Gene Clusters ◽  
Heterologous Proteins ◽  
Gene Encoding ◽  
Promising Alternative ◽  
High Yields ◽  
Melatonin Production

Abstract Background Melatonin has attracted substantial attention because of its excellent prospects for both medical applications and crop improvement. The microbial production of melatonin is a safer and more promising alternative to chemical synthesis approaches. Researchers have failed to produce high yields of melatonin in common heterologous hosts due to either the insolubility or low enzyme activity of proteins encoded by gene clusters related to melatonin biosynthesis. Results Here, a combinatorial gene pathway for melatonin production was successfully established in Escherichia coli by combining the physostigmine biosynthetic genes from Streptomyces albulus and gene encoding phenylalanine 4-hydroxylase (P4H) from Xanthomonas campestris and caffeic acid 3-O-methyltransferase (COMT) from Oryza sativa. A threefold improvement of melatonin production was achieved by balancing the expression of heterologous proteins and adding 3% glycerol. Further protein engineering and metabolic engineering were conducted to improve the conversion of N-acetylserotonin (NAS) to melatonin. Construction of COMT variant containing C303F and V321T mutations increased the production of melatonin by fivefold. Moreover, the deletion of speD gene increased the supply of S-adenosylmethionine (SAM), an indispensable cofactor of COMT, which doubled the yield of melatonin. In the final engineered strain EcMEL8, the production of NAS and melatonin reached 879.38 ± 71.42 mg/L and 136.17 ± 1.33 mg/L in a shake flask. Finally, in a 2-L bioreactor, EcMEL8 produced 1.06 ± 0.07 g/L NAS and 0.65 ± 0.11 g/L melatonin with tryptophan supplementation. Conclusions This study established a novel combinatorial pathway for melatonin biosynthesis in E. coli and provided alternative strategies for improvement of melatonin production.

scholarly journals Escherichia coli exonuclease VII. Cloning and sequencing of the gene encoding the large subunit (xseA).

1986 ◽  
Vol 261 (32) ◽  
pp. 14929-14935
Author(s):  
J W Chase ◽  
B A Rabin ◽  
J B Murphy ◽  
K L Stone ◽  
K R Williams
Keyword(s):  
Escherichia Coli ◽  
Large Subunit ◽  
Cloning And Sequencing ◽  
Gene Encoding ◽  
Exonuclease Vii

scholarly journals Identification of the Gene Encoding Sulfopyruvate Decarboxylase, an Enzyme Involved in Biosynthesis of Coenzyme M

2000 ◽  
Vol 182 (17) ◽  
pp. 4862-4867 ◽  
Author(s):  
Marion Graupner ◽  
Huimin Xu ◽  
Robert H. White
Keyword(s):  
Escherichia Coli ◽  
Second Step ◽  
Gene Products ◽  
Methanococcus Jannaschii ◽  
Coenzyme M ◽  
Gene Encoding ◽  
Fourth Step

ABSTRACT The products of two adjacent genes in the chromosome ofMethanococcus jannaschii are similar to the amino and carboxyl halves of phosphonopyruvate decarboxylase, the enzyme that catalyzes the second step of fosfomycin biosynthesis inStreptomyces wedmorensis. These two M. jannaschii genes were recombinantly expressed inEscherichia coli, and their gene products were tested for the ability to catalyze the decarboxylation of a series of α-ketoacids. Both subunits are required to form an α6β6 dodecamer that specifically catalyzes the decarboxylation of sulfopyruvic acid to sulfoacetaldehyde. This transformation is the fourth step in the biosynthesis of coenzyme M, a crucial cofactor in methanogenesis and aliphatic alkene metabolism. The M. jannaschiisulfopyruvate decarboxylase was found to be inactivated by oxygen and reactivated by reduction with dithionite. The two subunits, designated ComD and ComE, comprise the first enzyme for the biosynthesis of coenzyme M to be described.

Metabolic engineering of Escherichia coli for the production of malic acid

2008 ◽  
Vol 40 (2) ◽  
pp. 312-320 ◽  
Author(s):  
Soo Yun Moon ◽  
Soon Ho Hong ◽  
Tae Yong Kim ◽  
Sang Yup Lee
Keyword(s):  
Escherichia Coli ◽  
Metabolic Engineering ◽  
Malic Acid

Metabolic engineering and synthetic biology approaches driving isoprenoid production in Escherichia coli

2017 ◽  
Vol 241 ◽  
pp. 430-438 ◽  
Author(s):  
Chonglong Wang ◽  
Bakht Zada ◽  
Gongyuan Wei ◽  
Seon-Won Kim
Keyword(s):  
Escherichia Coli ◽  
Metabolic Engineering ◽  
Synthetic Biology
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