scholarly journals One-step biosynthesis of α-ketoisocaproate from l-leucine by an Escherichia coli whole-cell biocatalyst expressing an l-amino acid deaminase from Proteus vulgaris

2015 ◽  
Vol 5 (1) ◽  
Author(s):  
Yang Song ◽  
Jianghua Li ◽  
Hyun-dong Shin ◽  
Guocheng Du ◽  
Long Liu ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Amino Acid ◽  
Proteus Vulgaris ◽  
Whole Cell ◽  
Whole Cell Biocatalyst ◽  
One Step

Transporter engineering and enzyme evolution for pyruvate production from d/l-alanine with a whole-cell biocatalyst expressing l-amino acid deaminase from Proteus mirabilis

RSC Advances ◽  
2016 ◽  
Vol 6 (86) ◽  
pp. 82676-82684 ◽  
Author(s):  
Gazi Sakir Hossain ◽  
Hyun-dong Shin ◽  
Jianghua Li ◽  
Guocheng Du ◽  
Jian Chen ◽  
...  
Keyword(s):  
Amino Acid ◽  
Proteus Mirabilis ◽  
Production Method ◽  
Enzyme Evolution ◽  
Whole Cell ◽  
E Coli ◽  
Whole Cell Biocatalyst ◽  
One Step

Pyruvate, which has been widely used in the food, pharmaceutical, and agrochemical industries, can be produced by “one-step pyruvate production” method from d/l-alanine with a whole-cell E. coli biocatalyst expressing l-amino acid deaminase (pm1) from Proteus mirabilis.

scholarly journals Tuning a bi-enzymatic cascade reaction in Escherichia coli to facilitate NADPH regeneration for ε-caprolactone production

2021 ◽  
Vol 8 (1) ◽  
Author(s):  
Jinghui Xiong ◽  
Hefeng Chen ◽  
Ran Liu ◽  
Hao Yu ◽  
Min Zhuo ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Binding Sites ◽  
Regeneration System ◽  
Nadph Regeneration ◽  
Whole Cell ◽  
Cyclohexanone Monooxygenase ◽  
Ribosome Binding ◽  
Ribosome Binding Sites ◽  
Whole Cell Biocatalyst ◽  
Substrate Tolerance

Abstractε-Caprolactone is a monomer of poly(ε-caprolactone) which has been widely used in tissue engineering due to its biodegradability and biocompatibility. To meet the massive demand for this monomer, an efficient whole-cell biocatalytic approach was constructed to boost the ε-caprolactone production using cyclohexanol as substrate. Combining an alcohol dehydrogenase (ADH) with a cyclohexanone monooxygenase (CHMO) in Escherichia coli, a self-sufficient NADPH-cofactor regeneration system was obtained. Furthermore, some improved variants with the better substrate tolerance and higher catalytic ability to ε-caprolactone production were designed by regulating the ribosome binding sites. The best mutant strain exhibited an ε-caprolactone yield of 0.80 mol/mol using 60 mM cyclohexanol as substrate, while the starting strain only got a conversion of 0.38 mol/mol when 20 mM cyclohexanol was supplemented. The engineered whole-cell biocatalyst was used in four sequential batches to achieve a production of 126 mM ε-caprolactone with a high molar yield of 0.78 mol/mol.

Permeabilizing Escherichia coli for whole cell biocatalyst with enhanced biotransformation ability from l-glutamate to GABA

2014 ◽  
Vol 107 ◽  
pp. 39-46 ◽  
Author(s):  
Wei-rui Zhao ◽  
Jun Huang ◽  
Chun-long Peng ◽  
Sheng Hu ◽  
Pi-yu Ke ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Whole Cell ◽  
Whole Cell Biocatalyst

scholarly journals High-Level Conversion of l-lysine into Cadaverine by Escherichia coli Whole Cell Biocatalyst Expressing Hafnia alvei l-lysine Decarboxylase

Polymers ◽  
2019 ◽  
Vol 11 (7) ◽  
pp. 1184 ◽  
Author(s):  
Kim ◽  
Baritugo ◽  
Oh ◽  
Kang ◽  
Jung ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Protein Expression ◽  
Pyridoxal Phosphate ◽  
Lysine Decarboxylase ◽  
Hafnia Alvei ◽  
Whole Cell ◽  
E Coli ◽  
Whole Cell Biocatalyst ◽  
High Level ◽  
Whole Cell Bioconversion

Cadaverine is a C5 diamine monomer used for the production of bio-based polyamide 510. Cadaverine is produced by the decarboxylation of l-lysine using a lysine decarboxylase (LDC). In this study, we developed recombinant Escherichia coli strains for the expression of LDC from Hafnia alvei. The resulting recombinant XBHaLDC strain was used as a whole cell biocatalyst for the high-level bioconversion of l-lysine into cadaverine without the supplementation of isopropyl β-d-1-thiogalactopyranoside (IPTG) for the induction of protein expression and pyridoxal phosphate (PLP), a key cofactor for an LDC reaction. The comparison of results from enzyme characterization of E. coli and H. alvei LDC revealed that H. alvei LDC exhibited greater bioconversion ability than E. coli LDC due to higher levels of protein expression in all cellular fractions and a higher specific activity at 37 °C (1825 U/mg protein > 1003 U/mg protein). The recombinant XBHaLDC and XBEcLDC strains were constructed for the high-level production of cadaverine. Recombinant XBHaLDC produced a 1.3-fold higher titer of cadaverine (6.1 g/L) than the XBEcLDC strain (4.8 g/L) from 10 g/L of l-lysine. Furthermore, XBHaLDC, concentrated to an optical density (OD600) of 50, efficiently produced 136 g/L of cadaverine from 200 g/L of l-lysine (97% molar yield) via an IPTG- and PLP-free whole cell bioconversion reaction. Cadaverine synthesized via a whole cell biocatalyst reaction using XBHaLDC was purified to polymer grade, and purified cadaverine was successfully used for the synthesis of polyamide 510. In conclusion, an IPTG- and PLP-free whole cell bioconversion process of l-lysine into cadaverine, using recombinant XBHaLDC, was successfully utilized for the production of bio-based polyamide 510, which has physical and thermal properties similar to polyamide 510 synthesized from chemical-grade cadaverine.

scholarly journals Recovery of active beta-lactamases from Proteus vulgaris and RTEM-1 hybrid by random mutagenesis by using a dnaQ strain of Escherichia coli.

1996 ◽  
Vol 40 (9) ◽  
pp. 2152-2159 ◽  
Author(s):  
S M Hosseini-Mazinani ◽  
E Nakajima ◽  
Y Ihara ◽  
K Z Kameyama ◽  
K Sugimoto
Keyword(s):  
Escherichia Coli ◽  
Amino Acid ◽  
Random Mutagenesis ◽  
Amino Acid Residues ◽  
Proteus Vulgaris ◽  
Coding Region ◽  
Beta Lactamases ◽  
Functional Roles ◽  
Hybrid Genes ◽  
Lactamase Gene

Proteus vulgaris and RTEM-1 beta-lactamases that belong to molecular class A with 37% amino acid similarity were examined to find the relationship between amino acid residues and activity of enzymes. MICs of ampicillin were > 2,000 micrograms/ml for Escherichia coli cells producing these enzymes. We have made 18 hybrid genes by substituting the coding region of the P. vulgaris beta-lactamase gene with the equivalent portions from the RTEM-1 gene. Most of these hybrids produced inactive proteins, but a few hybrid enzymes had partial or trace activity. From one of the hybrid genes (MIC of ampicillin, 100 micrograms/ml), we recovered three kinds of active mutants which provided ampicillin MICs of 1,000 micrograms/ml by the selection of spontaneous mutations in a dnaQ strain of E. coli. In these mutants, Leu-148, Met-182, and Tyr-274 were replaced with Val, Thr, and His, respectively. These amino acids have not been identified as residues with functional roles in substrate hydrolysis. Furthermore, from these hybrid mutants, we obtained a second set of mutants which conferred ampicillin MICs of 1,500 micrograms/ml. Interestingly, the second mutations were limited to these three amino acid substitutions. These amino acid residues which do not directly interact with substrates have an effect on enzyme activity. These mutant enzymes exhibited lower K(m) values for cephaloridine than both parental enzymes.

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