Systematic engineering of transport and transcription to boost alkaline α-amylase production in Bacillus subtilis
In the present work, we used systematic engineering at transport and transcription levels to significantly enhance alkaline α-amylase production in Bacillus subtilis 168 M . Signal peptide YwbN’ proved to be optimal. Alkaline α-amylase production was elevated by deleting a putative peptide segment o...
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| Veröffentlicht in: | Applied microbiology and biotechnology 2020-04, Vol.104 (7), p.2973-2985 |
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| creator | Yang, Haiquan Ma, Yingfang Zhao, Yuan Shen, Wei Chen, Xianzhong |
| description | In the present work, we used systematic engineering at transport and transcription levels to significantly enhance alkaline α-amylase production in
Bacillus subtilis
168
M
. Signal peptide YwbN’ proved to be optimal. Alkaline α-amylase production was elevated by deleting a putative peptide segment of YwbN’. Insertion of arginine (R) between residues 5 and 6 of YwbN’∆p further increased the protein yield. Enhancing positive charges at sites 4 and 10 and decreasing the hydrophobicity of the
H
-region of YwbN’∆p were critical for improving alkaline α-amylase production in
B
.
subtilis
168
M
. P
HpaII
was the optimal promoter, and deleting − 27T or − 31A from P
HpaII
enhanced the transcription of the target gene. Using a single-pulse feeding-based fed-batch system, alkaline α-amylase activity of
B
.
subtilis
168
M
P
∆−27T
was increased by 250.6-fold, compared with
B
.
subtilis
168
M
A1. |
| doi_str_mv | 10.1007/s00253-020-10435-z |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_2353588297</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>2353588297</sourcerecordid><originalsourceid>FETCH-LOGICAL-c375t-8169ae783307ff8362fda209c55806569c6bb1b9c68a14591896468bfcbb2b003</originalsourceid><addsrcrecordid>eNp9kc1u1TAQhS0EopcLL8ACWWLDxjC2Y8dZQsWfVIlFy9qyfZ3KxbGDnSxu34oX6TPhNgUkFl2NRvOdM6M5CL2k8JYC9O8qABOcAANCoeOCXD9CO9pxRkDS7jHaAe0F6cWgTtCzWq8AKFNSPkUnnDWeKrVD5fxYFz-ZJTjs02VI3peQLnEe8VJMqnMuCzbpsHWuhHkJOeElY5tzbaP4w8Smwje_iJmO0VSP55IPq7vjQsIfjAsxrhXX1S4hhvocPRlNrP7Ffd2j758-Xpx-IWffPn89fX9GHO_FQhSVg_G94hz6cVRcsvFgGAxOCAVSyMFJa6ltRRnaiYGqQXZS2dFZyywA36M3m2-75-fq66KnUJ2P0SSf16oZF1woxYa-oa__Q6_yWlK7rlF9B4Po21_3iG2UK7nW4kc9lzCZctQU9G0iektEt0T0XSL6uole3VuvdvKHv5I_ETSAb0Cdbz_vy7_dD9j-BoAcmMM</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>2374095714</pqid></control><display><type>article</type><title>Systematic engineering of transport and transcription to boost alkaline α-amylase production in Bacillus subtilis</title><source>SpringerLink Journals - AutoHoldings</source><creator>Yang, Haiquan ; Ma, Yingfang ; Zhao, Yuan ; Shen, Wei ; Chen, Xianzhong</creator><creatorcontrib>Yang, Haiquan ; Ma, Yingfang ; Zhao, Yuan ; Shen, Wei ; Chen, Xianzhong</creatorcontrib><description>In the present work, we used systematic engineering at transport and transcription levels to significantly enhance alkaline α-amylase production in
Bacillus subtilis
168
M
. Signal peptide YwbN’ proved to be optimal. Alkaline α-amylase production was elevated by deleting a putative peptide segment of YwbN’. Insertion of arginine (R) between residues 5 and 6 of YwbN’∆p further increased the protein yield. Enhancing positive charges at sites 4 and 10 and decreasing the hydrophobicity of the
H
-region of YwbN’∆p were critical for improving alkaline α-amylase production in
B
.
subtilis
168
M
. P
HpaII
was the optimal promoter, and deleting − 27T or − 31A from P
HpaII
enhanced the transcription of the target gene. Using a single-pulse feeding-based fed-batch system, alkaline α-amylase activity of
B
.
subtilis
168
M
P
∆−27T
was increased by 250.6-fold, compared with
B
.
subtilis
168
M
A1.</description><identifier>ISSN: 0175-7598</identifier><identifier>EISSN: 1432-0614</identifier><identifier>DOI: 10.1007/s00253-020-10435-z</identifier><identifier>PMID: 32043188</identifier><language>eng</language><publisher>Berlin/Heidelberg: Springer Berlin Heidelberg</publisher><subject>Amylases ; Arginine ; Bacillus subtilis ; Batch culture ; Biomedical and Life Sciences ; Biotechnologically Relevant Enzymes and Proteins ; Biotechnology ; Hydrophobicity ; Life Sciences ; Microbial Genetics and Genomics ; Microbiology ; Peptides ; Transcription ; Transport ; α-Amylase</subject><ispartof>Applied microbiology and biotechnology, 2020-04, Vol.104 (7), p.2973-2985</ispartof><rights>Springer-Verlag GmbH Germany, part of Springer Nature 2020</rights><rights>Applied Microbiology and Biotechnology is a copyright of Springer, (2020). All Rights Reserved.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c375t-8169ae783307ff8362fda209c55806569c6bb1b9c68a14591896468bfcbb2b003</citedby><cites>FETCH-LOGICAL-c375t-8169ae783307ff8362fda209c55806569c6bb1b9c68a14591896468bfcbb2b003</cites><orcidid>0000-0001-6173-8787</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1007/s00253-020-10435-z$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1007/s00253-020-10435-z$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>314,780,784,27924,27925,41488,42557,51319</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/32043188$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Yang, Haiquan</creatorcontrib><creatorcontrib>Ma, Yingfang</creatorcontrib><creatorcontrib>Zhao, Yuan</creatorcontrib><creatorcontrib>Shen, Wei</creatorcontrib><creatorcontrib>Chen, Xianzhong</creatorcontrib><title>Systematic engineering of transport and transcription to boost alkaline α-amylase production in Bacillus subtilis</title><title>Applied microbiology and biotechnology</title><addtitle>Appl Microbiol Biotechnol</addtitle><addtitle>Appl Microbiol Biotechnol</addtitle><description>In the present work, we used systematic engineering at transport and transcription levels to significantly enhance alkaline α-amylase production in
Bacillus subtilis
168
M
. Signal peptide YwbN’ proved to be optimal. Alkaline α-amylase production was elevated by deleting a putative peptide segment of YwbN’. Insertion of arginine (R) between residues 5 and 6 of YwbN’∆p further increased the protein yield. Enhancing positive charges at sites 4 and 10 and decreasing the hydrophobicity of the
H
-region of YwbN’∆p were critical for improving alkaline α-amylase production in
B
.
subtilis
168
M
. P
HpaII
was the optimal promoter, and deleting − 27T or − 31A from P
HpaII
enhanced the transcription of the target gene. Using a single-pulse feeding-based fed-batch system, alkaline α-amylase activity of
B
.
subtilis
168
M
P
∆−27T
was increased by 250.6-fold, compared with
B
.
subtilis
168
M
A1.</description><subject>Amylases</subject><subject>Arginine</subject><subject>Bacillus subtilis</subject><subject>Batch culture</subject><subject>Biomedical and Life Sciences</subject><subject>Biotechnologically Relevant Enzymes and Proteins</subject><subject>Biotechnology</subject><subject>Hydrophobicity</subject><subject>Life Sciences</subject><subject>Microbial Genetics and Genomics</subject><subject>Microbiology</subject><subject>Peptides</subject><subject>Transcription</subject><subject>Transport</subject><subject>α-Amylase</subject><issn>0175-7598</issn><issn>1432-0614</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2020</creationdate><recordtype>article</recordtype><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNp9kc1u1TAQhS0EopcLL8ACWWLDxjC2Y8dZQsWfVIlFy9qyfZ3KxbGDnSxu34oX6TPhNgUkFl2NRvOdM6M5CL2k8JYC9O8qABOcAANCoeOCXD9CO9pxRkDS7jHaAe0F6cWgTtCzWq8AKFNSPkUnnDWeKrVD5fxYFz-ZJTjs02VI3peQLnEe8VJMqnMuCzbpsHWuhHkJOeElY5tzbaP4w8Smwje_iJmO0VSP55IPq7vjQsIfjAsxrhXX1S4hhvocPRlNrP7Ffd2j758-Xpx-IWffPn89fX9GHO_FQhSVg_G94hz6cVRcsvFgGAxOCAVSyMFJa6ltRRnaiYGqQXZS2dFZyywA36M3m2-75-fq66KnUJ2P0SSf16oZF1woxYa-oa__Q6_yWlK7rlF9B4Po21_3iG2UK7nW4kc9lzCZctQU9G0iektEt0T0XSL6uole3VuvdvKHv5I_ETSAb0Cdbz_vy7_dD9j-BoAcmMM</recordid><startdate>20200401</startdate><enddate>20200401</enddate><creator>Yang, 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Genomics</topic><topic>Microbiology</topic><topic>Peptides</topic><topic>Transcription</topic><topic>Transport</topic><topic>α-Amylase</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Yang, Haiquan</creatorcontrib><creatorcontrib>Ma, Yingfang</creatorcontrib><creatorcontrib>Zhao, Yuan</creatorcontrib><creatorcontrib>Shen, Wei</creatorcontrib><creatorcontrib>Chen, Xianzhong</creatorcontrib><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>ABI/INFORM Collection</collection><collection>ABI/INFORM Global (PDF only)</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>ABI/INFORM Global (Alumni 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Xianzhong</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Systematic engineering of transport and transcription to boost alkaline α-amylase production in Bacillus subtilis</atitle><jtitle>Applied microbiology and biotechnology</jtitle><stitle>Appl Microbiol Biotechnol</stitle><addtitle>Appl Microbiol Biotechnol</addtitle><date>2020-04-01</date><risdate>2020</risdate><volume>104</volume><issue>7</issue><spage>2973</spage><epage>2985</epage><pages>2973-2985</pages><issn>0175-7598</issn><eissn>1432-0614</eissn><abstract>In the present work, we used systematic engineering at transport and transcription levels to significantly enhance alkaline α-amylase production in
Bacillus subtilis
168
M
. Signal peptide YwbN’ proved to be optimal. Alkaline α-amylase production was elevated by deleting a putative peptide segment of YwbN’. Insertion of arginine (R) between residues 5 and 6 of YwbN’∆p further increased the protein yield. Enhancing positive charges at sites 4 and 10 and decreasing the hydrophobicity of the
H
-region of YwbN’∆p were critical for improving alkaline α-amylase production in
B
.
subtilis
168
M
. P
HpaII
was the optimal promoter, and deleting − 27T or − 31A from P
HpaII
enhanced the transcription of the target gene. Using a single-pulse feeding-based fed-batch system, alkaline α-amylase activity of
B
.
subtilis
168
M
P
∆−27T
was increased by 250.6-fold, compared with
B
.
subtilis
168
M
A1.</abstract><cop>Berlin/Heidelberg</cop><pub>Springer Berlin Heidelberg</pub><pmid>32043188</pmid><doi>10.1007/s00253-020-10435-z</doi><tpages>13</tpages><orcidid>https://orcid.org/0000-0001-6173-8787</orcidid></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0175-7598 |
| ispartof | Applied microbiology and biotechnology, 2020-04, Vol.104 (7), p.2973-2985 |
| issn | 0175-7598 1432-0614 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_2353588297 |
| source | SpringerLink Journals - AutoHoldings |
| subjects | Amylases Arginine Bacillus subtilis Batch culture Biomedical and Life Sciences Biotechnologically Relevant Enzymes and Proteins Biotechnology Hydrophobicity Life Sciences Microbial Genetics and Genomics Microbiology Peptides Transcription Transport α-Amylase |
| title | Systematic engineering of transport and transcription to boost alkaline α-amylase production in Bacillus subtilis |
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