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.

Bioconversion of l-glutamic acid to α-ketoglutaric acid by an immobilized whole-cell biocatalyst expressing l-amino acid deaminase from Proteus mirabilis

2014 ◽  
Vol 169 ◽  
pp. 112-120 ◽  
Author(s):  
Gazi Sakir Hossain ◽  
Jianghua Li ◽  
Hyun-dong Shin ◽  
Rachel R. Chen ◽  
Guocheng Du ◽  
...  
Keyword(s):  
Amino Acid ◽  
Glutamic Acid ◽  
Proteus Mirabilis ◽  
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.

Isolation of Labeled Lipoprotein from Escherichia coli and Proteus mirabilis after Incubation with [14C]Penicillin

1985 ◽  
Vol 40 (5-6) ◽  
pp. 415-420 ◽  
Author(s):  
Gerhard Gruner ◽  
Monier H. Tadros ◽  
Roland Plapp
Keyword(s):  
Escherichia Coli ◽  
Amino Acid ◽  
Amino Acid Composition ◽  
Acid Composition ◽  
Proteus Mirabilis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Apparent Molecular Weight ◽  
Penicillin G ◽  
Free Form ◽  
E Coli

Abstract [14C]penicillin binding experiments and membrane analysis were carried out with cell envelope preparations from Escherichia coli and Proteus mirabilis. After incubation with [14C] penicillin G labeled free lipoprotein could be identified. The analysis of the isolated lipoprotein by SDS polyacrylamide gel electrophoresis indicates that there is only one protein with an apparent molecular weight of 7000. The amino acid composition of isolated labeled free lipoprotein from E. coli was identical to the lipoprotein already found in E. coli. It is a point of interest that the amino acid composition of the isolated labeled free lipoprotein from P. mirabilis D52 differs from that found in other mutants of this strain. The free form of lipoprotein from P. mirabilis D52 is composed of 61 amino acids and has glycine, phenylalanine and proline as specific components.

scholarly journals Short-Chain Flavor Ester Synthesis in Organic Media by an E. coli Whole-Cell Biocatalyst Expressing a Newly Characterized Heterologous Lipase

PLoS ONE ◽  
2014 ◽  
Vol 9 (3) ◽  
pp. e91872 ◽  
Author(s):  
Guillaume Brault ◽  
François Shareck ◽  
Yves Hurtubise ◽  
François Lépine ◽  
Nicolas Doucet
Keyword(s):  
Short Chain ◽  
Organic Media ◽  
Ester Synthesis ◽  
Whole Cell ◽  
E Coli ◽  
Whole Cell Biocatalyst ◽  
Flavor Ester Synthesis

scholarly journals New Plasmid-Mediated Quinolone Resistance Gene, qnrC, Found in a Clinical Isolate of Proteus mirabilis

2009 ◽  
Vol 53 (5) ◽  
pp. 1892-1897 ◽  
Author(s):  
Minghua Wang ◽  
Qinglan Guo ◽  
Xiaogang Xu ◽  
Xiaoying Wang ◽  
Xinyu Ye ◽  
...  
Keyword(s):  
Amino Acid ◽  
Resistance Gene ◽  
Clinical Isolate ◽  
Proteus Mirabilis ◽  
Amino Acid Identity ◽  
Quinolone Resistance ◽  
Clinical Strain ◽  
E Coli ◽  
Acid Protein ◽  
New Gene

ABSTRACT Since the discovery of qnrA in 1998, two additional qnr genes, qnrB and qnrS, have been described. These three plasmid-mediated genes contribute to quinolone resistance in gram-negative pathogens worldwide. A clinical strain of Proteus mirabilis was isolated from an outpatient with a urinary tract infection and was susceptible to most antimicrobials but resistant to ampicillin, sulfamethoxazole, and trimethoprim. Plasmid pHS10, harbored by this strain, was transferred to azide-resistant Escherichia coli J53 by conjugation. A transconjugant with pHS10 had low-level quinolone resistance but was negative by PCR for the known qnr genes, aac(6′)-Ib-cr and qepA. The ciprofloxacin MIC for the clinical strain and a J53/pHS10 transconjugant was 0.25 μg/ml, representing an increase of 32-fold relative to that for the recipient, J53. The plasmid was digested with HindIII, and a 4.4-kb DNA fragment containing the new gene was cloned into pUC18 and transformed into E. coli TOP10. Sequencing showed that the responsible 666-bp gene, designated qnrC, encoded a 221-amino-acid protein, QnrC, which shared 64%, 42%, 59%, and 43% amino acid identity with QnrA1, QnrB1, QnrS1, and QnrD, respectively. Upstream of qnrC there existed a new IS3 family insertion sequence, ISPmi1, which encoded a frameshifted transposase. qnrC could not be detected by PCR, however, in 2,020 strains of Enterobacteriaceae. A new quinolone resistance gene, qnrC, was thus characterized from plasmid pHS10 carried by a clinical isolate of P. mirabilis.

scholarly journals Development of an Autofluorescent Whole-Cell Biocatalyst by Displaying Dual Functional Moieties on Escherichia coli Cell Surfaces and Construction of a Coculture with Organophosphate-Mineralizing Activity

2008 ◽  
Vol 74 (24) ◽  
pp. 7733-7739 ◽  
Author(s):  
Chao Yang ◽  
Yaran Zhu ◽  
Jijian Yang ◽  
Zheng Liu ◽  
Chuanling Qiao ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Fluorescent Protein ◽  
Surface Display ◽  
Ice Nucleation Protein ◽  
Whole Cell ◽  
E Coli ◽  
Dual Functional ◽  
Whole Cell Biocatalyst ◽  
Mineralizing Activity ◽  
Green Fluorescent

ABSTRACT Surface display of the active proteins on living cells has enormous potential in the degradation of numerous toxic compounds. Here, we report the codisplay of organophosphorus hydrolase (OPH) and enhanced green fluorescent protein (GFP) on the cell surface of Escherichia coli by use of the truncated ice nucleation protein (INPNC) and Lpp-OmpA fusion systems. The surface localization of both INPNC-OPH and Lpp-OmpA-GFP was demonstrated by Western blot analysis, immunofluorescence microscopy, and a protease accessibility experiment. Anchorage of GFP and OPH on the outer membrane neither inhibits cell growth nor affects cell viability, as shown by growth kinetics of cells and stability of resting cultures. The engineered E. coli can be applied in the form of a whole-cell biocatalyst and can be tracked by fluorescence during bioremediation. This strategy of codisplay should open a new dimension for the display of multiple functional moieties on the surface of a bacterial cell. Furthermore, a coculture comprised of the engineered E. coli and a natural p-nitrophenol (PNP) degrader, Ochrobactrum sp. strain LL-1, was assembled for complete mineralization of organophosphates (OPs) with a PNP substitution. The coculture degraded OPs as well as PNP rapidly. Therefore, the coculture with autofluorescent and mineralizing activities can potentially be applied for bioremediation of OP-contaminated sites.

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