Revision as of 15:28, 12 October 2022 (view source) Taaa (Talk | contribs) ← Older edit		Latest revision as of 17:15, 13 October 2022 (view source) Kuaikuai (Talk | contribs)
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	<partinfo>BBa_K4325003 short</partinfo>		<partinfo>BBa_K4325003 short</partinfo>
	===Description===		===Description===
−	<p>gshF encodes a bifunctional glutathione synthetase GshF originated from Streptococcus thermophilus.</p>	+	<p><i>gshF</i> encodes a bifunctional glutathione synthetase GshF originated from <i>Streptococcus thermophilus</i>.</p>
	===Usage===		===Usage===
	<p>It is less sensitive to GSH, is and codon optimized for expressing in <i>G. hansenii</i>.		<p>It is less sensitive to GSH, is and codon optimized for expressing in <i>G. hansenii</i>.
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	=2022 SZPT-China=		=2022 SZPT-China=
−	<h3>Characterization</h3>	+	<h3>1.Characterization in <i>E. coli</i>  TOP10</h3>
−	<h4>	+	As shown in Figure 2,  the expression of GshF which encoded by BBa_K4325003 were verified successfully by PCR amplification and western blot respectively. The GSH production of <i>E. coli</i> TOP10 containing this part is much higher (~92 fold) than that of wild-type <i>E. coli</i> TOP10.</p>
−	1. Verification of Western blot electrophoresis in <i> E. coli</i>.</h4>	+	[[File:K15 2.png|800px|thumb|center|Figure 2: (a) Verification of gshF in <i>E. coli</i> TOP10; (b) Verification of western blot electrophoresis in <i>E. coli</i> Top10; (c) Comparison of GSH production between wild type and engineered of <i>E. coli</i> Top10.]]
−	[[File:Mfold-K3257013-1.png|600px|thumb|center|Figure1: Verification of Western blot electrophoresis in <i> E. coli.</i>]]	+	<br>
−	As shown in Figure 1, the expression of GshF were verified successfully by Western blot.</p>	+	<h3>2.Characterization in <i>G. hansenii</i>  ATCC53582</h3>
−	<h4>2.References</h4>	+	<p>Figure3 (a) shows the DNA fragments of gshF were amplified from <i>G. hansenii</i>, thus confirming the successful incorporation of the plasmid. Figure2 (b) shows that <i>G. hansenii</i> containing this part exhibited enhanced GSH biosynthesis, as evidenced by colorimetric analysis of glutathione.</p>
−	<p>[1] Li, W., Li, Z., Yang, J. & Ye, Q. Production of glutathione using a bifunctional enzyme encoded by gshF from Streptococcus thermophilus expressed in <i>Escherichia coli</i>. J. Biotechnol. <b>154</b>, 261-268 (2011).</p>	+
−	<p>[2] Pophaly, S. D. et al. Glutathione biosynthesis and activity of dependent enzymes in food-grade lactic acid bacteria harbouring multidomain bifunctional fusion gene (gshF). J. Appl. Microbiol. <b>123</b>, 194-203 (2017).</p>	+	[[File:K15 3.png|600px|thumb|center|Figure 3: (a) Verification of gshF in <i>G. hansenii</i> ATCC53582; (b) Comparison of GSH production between wild type and engineered <i>G. hansenii</i>.]]
		+	<h3>References</h3>
		+	<p>[1] Li, W., Li, Z., Yang, J. & Ye, Q. Production of glutathione using a bifunctional enzyme encoded by gshF from Streptococcus thermophilus expressed in Escherichia coli. J. Biotechnol. <b>154</b>, 261-268 (2011.</p>
		+	<p>[2] Wang, D., Wang, C., Wu, H., Li, Z. & Ye, Q. Glutathione production by recombinant Escherichia coli expressing bifunctional glutathione synthetase. J. Ind. Microbiol. Biotechnol. <b>43</b>, 45-53 (2016).</p>
		+	<p>[3] Pophaly, S. D. et al. Glutathione biosynthesis and activity of dependent enzymes in food-grade lactic acid bacteria harbouring multidomain bifunctional fusion gene (gshF). J. Appl. Microbiol. <b>123</b>, 194-203 (2017).</p>
		+	<p>[4] Xiong, Z.-Q. et al. Functional analysis and heterologous expression of bifunctional glutathione synthetase from Lactobacillus. J. Dairy Sci.<b>101</b> , 6937-6945 (2018).</p>
		+	<p>[5] Cui, X. et al. Efficient glutathione production in metabolically engineered Escherichia coli strains using constitutive promoters. J. Biotechnol.<b>289</b>, 39-45 (2019).</p>

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Latest revision as of 17:15, 13 October 2022

gshF

Description

gshF encodes a bifunctional glutathione synthetase GshF originated from Streptococcus thermophilus.

Usage

It is less sensitive to GSH, is and codon optimized for expressing in G. hansenii.

Sequence and Features

2022 SZPT-China

1.Characterization in E. coli TOP10

As shown in Figure 2, the expression of GshF which encoded by BBa_K4325003 were verified successfully by PCR amplification and western blot respectively. The GSH production of E. coli TOP10 containing this part is much higher (~92 fold) than that of wild-type E. coli TOP10.

2.Characterization in G. hansenii ATCC53582

Figure3 (a) shows the DNA fragments of gshF were amplified from G. hansenii, thus confirming the successful incorporation of the plasmid. Figure2 (b) shows that G. hansenii containing this part exhibited enhanced GSH biosynthesis, as evidenced by colorimetric analysis of glutathione.

References

[1] Li, W., Li, Z., Yang, J. & Ye, Q. Production of glutathione using a bifunctional enzyme encoded by gshF from Streptococcus thermophilus expressed in Escherichia coli. J. Biotechnol. 154, 261-268 (2011.

[2] Wang, D., Wang, C., Wu, H., Li, Z. & Ye, Q. Glutathione production by recombinant Escherichia coli expressing bifunctional glutathione synthetase. J. Ind. Microbiol. Biotechnol. 43, 45-53 (2016).

[3] Pophaly, S. D. et al. Glutathione biosynthesis and activity of dependent enzymes in food-grade lactic acid bacteria harbouring multidomain bifunctional fusion gene (gshF). J. Appl. Microbiol. 123, 194-203 (2017).

[4] Xiong, Z.-Q. et al. Functional analysis and heterologous expression of bifunctional glutathione synthetase from Lactobacillus. J. Dairy Sci.101 , 6937-6945 (2018).

[5] Cui, X. et al. Efficient glutathione production in metabolically engineered Escherichia coli strains using constitutive promoters. J. Biotechnol.289, 39-45 (2019).