High-level production penicillin G acylase from Alcaligenes faecalis in recombinant Escherichia coli with optimization of carbon sources

Cheng, Shiwei; Song, Qing; Wei, Dongzhi; Gao, Bingxue

doi:10.1016/j.enzmictec.2007.02.011

Cited by 21 publications

(8 citation statements)

References 24 publications

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“…The amount of protein produced per gram dry cell weight (DCW) of T. kodakarensis was ϳ16 mg protein g DCW Ϫ1 with the ChiA⌬4 protein and is comparable to those of other microbial host cells. Protein production levels of 7 to over 150 mg protein g DCW Ϫ1 have been reported for E. coli (8,15,17), while 40 to over 140 mg protein g DCW Ϫ1 have been reported for Bacillus subtilis (36). …”

Section: Discussionmentioning

confidence: 99%

Thermococcus kodakarensis as a Host for Gene Expression and Protein Secretion

Takemasa

Yokooji

Yasuda

et al. 2011

Appl Environ Microbiol

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Taking advantage of the gene manipulation system developed in Thermococcus kodakarensis, here, we developed a system for gene expression and efficient protein secretion using this hyperthermophilic archaeon as a host cell. DNA fragments encoding the C-terminal domain of chitinase (ChiA⌬4), which exhibits endochitinase activity, and the putative signal sequence of a subtilisin-like protease (TK1675) were fused and positioned under the control of the strong constitutive promoter of the cell surface glycoprotein gene. This gene cassette was introduced into T. kodakarensis, and secretion of the ChiA⌬4 protein was examined. ChiA⌬4 was found exclusively in the culture supernatant and was not detected in the soluble and membrane fractions of the cell extract. The signal peptide was specifically cleaved at the C-terminal peptide bond following the Ala-Ser-Ala sequence. Efficient secretion of the orotidine-5-monophosphate decarboxylase protein was also achieved with the same strategy. We next individually overexpressed two genes (TK1675 and TK1689) encoding proteases with putative signal sequences. By comparing protein degradation activities in the host cells and transformants in both solid and liquid media, as well as measuring peptidase activity using synthetic peptide substrates, we observed dramatic increases in protein degradation activity in the two transformants. This study displays an initial demonstration of cell engineering in hyperthermophiles.

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Section: Discussionmentioning

confidence: 99%

Thermococcus kodakarensis as a Host for Gene Expression and Protein Secretion

Takemasa

Yokooji

Yasuda

et al. 2011

Appl Environ Microbiol

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“…Alcaligenes faecalis produces antibiotic compounds capable of inhibiting the growth of Gram-positive and Gram-negative bacteria (Zahir et al, 2013). Recombinant of A. faecalis with Escherichia coli capable of producing penicillin G in high quantities (Cheng et al, 2007). Biological control bacteria UPC 71 was 98% identity Pseudomonas stutzeri.…”

Section: Discussionmentioning

confidence: 99%

Isolation and Application of Pathogenic and Biological Control Bacteria from Pupae of Golden Silkworm Cricula trifenestrata Helfer

Uli¹,

Mubarik²,

Solihin³

2015

Research J. of Microbiology

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The decreasing productivity of silkworm cocoon is caused by attack of bacteria, fungi, protozoa and virus. The research was aimed to isolate of suspected pathogenic and biological control bacteria from damage and healthy pupae of golden silkworm (Cricula trifenestrata Helfer) and to do in vivo test of the bacteria toward 4th instar larvae of C. trifenestrata and mulberry silkworms, Bombyx mori Linn. The isolate UPC 60 was suspected as pathogenic bacteria due to cause the mortality of silkworms larvae. The bacteria UPC 40 and UPC 71 were selected as biological control bacteria which had capability to reduce the mortality of silkworms about 56.67 and 43.33%, respectively. Based on 16s RNA gene the bacteria UPC 40 had 97% identity with Alcaligenes faecalis, bacteria UPC 60 had 97% identity with Aeromonas dhakensis and bacteria UPC 71 had 98% identity with Pseudomonas stutzuri. The biological control bacteria showed antagonistic activity toward the pathogenic bacteria. The outcomes of the research that the biological control bacteria expected to be used to reduce pathogenic bacteria in the maintenance of silkworm so that the production and quality of silk yarn can be improved.

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“…Penicillin G acylase (PGA) can be synthesized by various microorganisms including bacteria, fungi and yeasts from a wide range of sources [1]. As an important biocatalyst [2] in the industrial production of β-lactam antibiotics, PGA could also catalyze the production of important semisynthetic intermediates of β-lactam antibiotics such as the mother nucleus 6-aminopenicillitic acid (6-APA), 7aminodeethyloxycephalosporic acid (7-ADCA) or 7aminocephalosporic acid (7-ACA) [3].…”

Section: Introductionmentioning

confidence: 99%