Molecular structure of cyclomaltodextrinase derived from amylolytic lactic acid bacterium Enterococcus faecium K-1 and properties of recombinant enzymes expressed in Escherichia coli and Lactobacillus plantarum

© 2017 Elsevier B.V. Gene encoding cyclomaltodextrinase (Cdx) from amylolytic lactic acid bacterium Enterococcus faecium K-1 was cloned and nucleotide sequence was analyzed. The open-reading frame consisted of 1767 bp encoding 588 deduced amino acids. Consequently, four typically conserved regions o...

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Main Authors: Kridsada Unban, Apinun Kanpiengjai, Saisamorn Lumyong, Thu Ha Nguyen, Dietmar Haltrich, Chartchai Khanongnuch
Format: Journal
Published: 2018
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spelling th-cmuir.6653943832-582982018-09-05T04:22:21Z Molecular structure of cyclomaltodextrinase derived from amylolytic lactic acid bacterium Enterococcus faecium K-1 and properties of recombinant enzymes expressed in Escherichia coli and Lactobacillus plantarum Kridsada Unban Apinun Kanpiengjai Saisamorn Lumyong Thu Ha Nguyen Dietmar Haltrich Chartchai Khanongnuch Biochemistry, Genetics and Molecular Biology © 2017 Elsevier B.V. Gene encoding cyclomaltodextrinase (Cdx) from amylolytic lactic acid bacterium Enterococcus faecium K-1 was cloned and nucleotide sequence was analyzed. The open-reading frame consisted of 1767 bp encoding 588 deduced amino acids. Consequently, four typically conserved regions of the glycoside hydrolase family 13 were revealed; however, nine exceeding amino acids (DSYQMTDVP) were found at the 282–290 position in comparison to previously reported cyclomaltodextrinases. This difference is believed to have an influence on the substrate specificity of this enzyme. The recombinant CDases expressed in Escherichia coli BL21 (CDX_E) and Lactobacillus plantarum WCFS1 (CDX_L) with high expression levels of 8041 and 5511 U/L were purified by Ni-NTA affinity chromatography. The active form CDX is a dimeric protein with two identical subunits of 62 kDa, approximately. Both CDX_E and CDX_L revealed nearly similar properties, but the thermostability of CDX_L was slightly higher. Mn2+and Co2+at a concentration of 1 mM stimulated the enzyme activity, while the Ag+, Cu2+and SDS solution completely inhibited enzyme activity. CDX exhibited the highest activity with α-cyclodextrin and β-cyclodextrin, but lower toward pullulan and starch. Importantly, this is the first report describing genes, the molecular structure and properties of cyclomaltodextrinase derived from lactic acid bacteria E. faecium. 2018-09-05T04:22:21Z 2018-09-05T04:22:21Z 2018-02-01 Journal 18790003 01418130 2-s2.0-85029773268 10.1016/j.ijbiomac.2017.09.060 https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=85029773268&origin=inward http://cmuir.cmu.ac.th/jspui/handle/6653943832/58298
institution Chiang Mai University
building Chiang Mai University Library
country Thailand
collection CMU Intellectual Repository
topic Biochemistry, Genetics and Molecular Biology
spellingShingle Biochemistry, Genetics and Molecular Biology
Kridsada Unban
Apinun Kanpiengjai
Saisamorn Lumyong
Thu Ha Nguyen
Dietmar Haltrich
Chartchai Khanongnuch
Molecular structure of cyclomaltodextrinase derived from amylolytic lactic acid bacterium Enterococcus faecium K-1 and properties of recombinant enzymes expressed in Escherichia coli and Lactobacillus plantarum
description © 2017 Elsevier B.V. Gene encoding cyclomaltodextrinase (Cdx) from amylolytic lactic acid bacterium Enterococcus faecium K-1 was cloned and nucleotide sequence was analyzed. The open-reading frame consisted of 1767 bp encoding 588 deduced amino acids. Consequently, four typically conserved regions of the glycoside hydrolase family 13 were revealed; however, nine exceeding amino acids (DSYQMTDVP) were found at the 282–290 position in comparison to previously reported cyclomaltodextrinases. This difference is believed to have an influence on the substrate specificity of this enzyme. The recombinant CDases expressed in Escherichia coli BL21 (CDX_E) and Lactobacillus plantarum WCFS1 (CDX_L) with high expression levels of 8041 and 5511 U/L were purified by Ni-NTA affinity chromatography. The active form CDX is a dimeric protein with two identical subunits of 62 kDa, approximately. Both CDX_E and CDX_L revealed nearly similar properties, but the thermostability of CDX_L was slightly higher. Mn2+and Co2+at a concentration of 1 mM stimulated the enzyme activity, while the Ag+, Cu2+and SDS solution completely inhibited enzyme activity. CDX exhibited the highest activity with α-cyclodextrin and β-cyclodextrin, but lower toward pullulan and starch. Importantly, this is the first report describing genes, the molecular structure and properties of cyclomaltodextrinase derived from lactic acid bacteria E. faecium.
format Journal
author Kridsada Unban
Apinun Kanpiengjai
Saisamorn Lumyong
Thu Ha Nguyen
Dietmar Haltrich
Chartchai Khanongnuch
author_facet Kridsada Unban
Apinun Kanpiengjai
Saisamorn Lumyong
Thu Ha Nguyen
Dietmar Haltrich
Chartchai Khanongnuch
author_sort Kridsada Unban
title Molecular structure of cyclomaltodextrinase derived from amylolytic lactic acid bacterium Enterococcus faecium K-1 and properties of recombinant enzymes expressed in Escherichia coli and Lactobacillus plantarum
title_short Molecular structure of cyclomaltodextrinase derived from amylolytic lactic acid bacterium Enterococcus faecium K-1 and properties of recombinant enzymes expressed in Escherichia coli and Lactobacillus plantarum
title_full Molecular structure of cyclomaltodextrinase derived from amylolytic lactic acid bacterium Enterococcus faecium K-1 and properties of recombinant enzymes expressed in Escherichia coli and Lactobacillus plantarum
title_fullStr Molecular structure of cyclomaltodextrinase derived from amylolytic lactic acid bacterium Enterococcus faecium K-1 and properties of recombinant enzymes expressed in Escherichia coli and Lactobacillus plantarum
title_full_unstemmed Molecular structure of cyclomaltodextrinase derived from amylolytic lactic acid bacterium Enterococcus faecium K-1 and properties of recombinant enzymes expressed in Escherichia coli and Lactobacillus plantarum
title_sort molecular structure of cyclomaltodextrinase derived from amylolytic lactic acid bacterium enterococcus faecium k-1 and properties of recombinant enzymes expressed in escherichia coli and lactobacillus plantarum
publishDate 2018
url https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=85029773268&origin=inward
http://cmuir.cmu.ac.th/jspui/handle/6653943832/58298
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