A new [beta]-galactosidase extracted from the infant feces with high hydrolytic and transgalactosylation activity
A [beta]-galactosidase ([beta]-Gal.sub.INF) was directly isolated from feces of an 8-month-old infant. Mass spectrum analysis showed [beta]-Gal.sub.INF with coverage over 50% to the [beta]-galactosidase from Bifidobacterium longum EK3. Accordingly, the [beta]-gal.sub.INF was amplified from the feces...
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| Veröffentlicht in: | Applied microbiology and biotechnology 2019-10, Vol.103 (20), p.8439 |
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| creator | Xin, Yongping Guo, Tingting Zhang, Yi Wu, Jiapeng Kong, Jian |
| description | A [beta]-galactosidase ([beta]-Gal.sub.INF) was directly isolated from feces of an 8-month-old infant. Mass spectrum analysis showed [beta]-Gal.sub.INF with coverage over 50% to the [beta]-galactosidase from Bifidobacterium longum EK3. Accordingly, the [beta]-gal.sub.INF was amplified from the feces metagenomic DNA by degenerate primers. After overexpressed in Escherichia coli, the [beta]-Gal.sub.INF was purified and biochemical characterized. [beta]-Gal.sub.INF existed as homotetramer and homodimer, whose activity (optimal at 50 °C, pH 6.5) was exhilaratingly increased to 484% by artificial intestinal juice. The K.sub.m and V.sub.max values for oNPG and lactose were 20.95 ± 2.76 mM, 5004.50 ± 318.8 [mu]mol min.sup.-1 mg.sup.-1 and 140.2 ± 17.7 mM, 293.1 ± 14.7 [mu]mol min.sup.-1 mg.sup.-1, respectively. The production rate of galacto-oligosaccharides by [beta]-Gal.sub.INF from 20% lactose at 50 °C was 33.4 ± 0.67%. These results suggested the [beta]-Gal.sub.INF with high hydrolytic and transgalactosylation activity from the infant intestinal has great potential as infant lactase preparation. Moreover, this study provided a new way for exploring undetected enzymes by uncultured-dependent methods. |
| doi_str_mv | 10.1007/s00253-019-10092-x |
| format | Article |
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Mass spectrum analysis showed [beta]-Gal.sub.INF with coverage over 50% to the [beta]-galactosidase from Bifidobacterium longum EK3. Accordingly, the [beta]-gal.sub.INF was amplified from the feces metagenomic DNA by degenerate primers. After overexpressed in Escherichia coli, the [beta]-Gal.sub.INF was purified and biochemical characterized. [beta]-Gal.sub.INF existed as homotetramer and homodimer, whose activity (optimal at 50 °C, pH 6.5) was exhilaratingly increased to 484% by artificial intestinal juice. The K.sub.m and V.sub.max values for oNPG and lactose were 20.95 ± 2.76 mM, 5004.50 ± 318.8 [mu]mol min.sup.-1 mg.sup.-1 and 140.2 ± 17.7 mM, 293.1 ± 14.7 [mu]mol min.sup.-1 mg.sup.-1, respectively. The production rate of galacto-oligosaccharides by [beta]-Gal.sub.INF from 20% lactose at 50 °C was 33.4 ± 0.67%. These results suggested the [beta]-Gal.sub.INF with high hydrolytic and transgalactosylation activity from the infant intestinal has great potential as infant lactase preparation. Moreover, this study provided a new way for exploring undetected enzymes by uncultured-dependent methods.</description><identifier>ISSN: 0175-7598</identifier><identifier>EISSN: 1432-0614</identifier><identifier>DOI: 10.1007/s00253-019-10092-x</identifier><language>eng</language><publisher>Springer</publisher><subject>Analysis ; Enzymes ; Escherichia coli ; Ethylenediaminetetraacetic acid ; Lactase ; Lactose</subject><ispartof>Applied microbiology and biotechnology, 2019-10, Vol.103 (20), p.8439</ispartof><rights>COPYRIGHT 2019 Springer</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,777,781,27905,27906</link.rule.ids></links><search><creatorcontrib>Xin, Yongping</creatorcontrib><creatorcontrib>Guo, Tingting</creatorcontrib><creatorcontrib>Zhang, Yi</creatorcontrib><creatorcontrib>Wu, Jiapeng</creatorcontrib><creatorcontrib>Kong, Jian</creatorcontrib><title>A new [beta]-galactosidase extracted from the infant feces with high hydrolytic and transgalactosylation activity</title><title>Applied microbiology and biotechnology</title><description>A [beta]-galactosidase ([beta]-Gal.sub.INF) was directly isolated from feces of an 8-month-old infant. Mass spectrum analysis showed [beta]-Gal.sub.INF with coverage over 50% to the [beta]-galactosidase from Bifidobacterium longum EK3. Accordingly, the [beta]-gal.sub.INF was amplified from the feces metagenomic DNA by degenerate primers. After overexpressed in Escherichia coli, the [beta]-Gal.sub.INF was purified and biochemical characterized. [beta]-Gal.sub.INF existed as homotetramer and homodimer, whose activity (optimal at 50 °C, pH 6.5) was exhilaratingly increased to 484% by artificial intestinal juice. The K.sub.m and V.sub.max values for oNPG and lactose were 20.95 ± 2.76 mM, 5004.50 ± 318.8 [mu]mol min.sup.-1 mg.sup.-1 and 140.2 ± 17.7 mM, 293.1 ± 14.7 [mu]mol min.sup.-1 mg.sup.-1, respectively. The production rate of galacto-oligosaccharides by [beta]-Gal.sub.INF from 20% lactose at 50 °C was 33.4 ± 0.67%. These results suggested the [beta]-Gal.sub.INF with high hydrolytic and transgalactosylation activity from the infant intestinal has great potential as infant lactase preparation. Moreover, this study provided a new way for exploring undetected enzymes by uncultured-dependent methods.</description><subject>Analysis</subject><subject>Enzymes</subject><subject>Escherichia coli</subject><subject>Ethylenediaminetetraacetic acid</subject><subject>Lactase</subject><subject>Lactose</subject><issn>0175-7598</issn><issn>1432-0614</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2019</creationdate><recordtype>article</recordtype><recordid>eNptkE9LAzEQxYMoWKtfwFPAk4fUSXY3yR5L0VooCP45iZTp7uw2st3FJtrutzeiggUZSPLC7z1mhrFzCSMJYK48gMoSATIXUedK7A7YQKaJEqBlesgGIE0mTJbbY3bi_SuAVFbrAXsb85a2_HlJAV9EjQ0WofOuRE-cdmETJZW82nRrHlbEXVthG3hFBXm-dWHFV66OR19uuqYPruDYljzaWv-b1TcYXNfyKNyHC_0pO6qw8XT2cw_Z08314-RWzO-ms8l4LmoJ0goiArNEbWyWQxGnIFMRgtIKU6lUhjLHNL4wKZLMVjpd6txqo9NUSQsIyZBdfOfGRmgRG---plk7XyzGGpIIKWMjNfqHilXS2hVdS5WL_3uGyz1DZEJcVI3v3i9mD_d_2U_7_Hq4</recordid><startdate>20191001</startdate><enddate>20191001</enddate><creator>Xin, Yongping</creator><creator>Guo, Tingting</creator><creator>Zhang, Yi</creator><creator>Wu, Jiapeng</creator><creator>Kong, Jian</creator><general>Springer</general><scope>ISR</scope></search><sort><creationdate>20191001</creationdate><title>A new [beta]-galactosidase extracted from the infant feces with high hydrolytic and transgalactosylation activity</title><author>Xin, Yongping ; Guo, Tingting ; Zhang, Yi ; Wu, Jiapeng ; Kong, Jian</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-g1018-eee07ba678590c061e7fea0262a41225a19a4412a3c358f64b698676442180a03</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2019</creationdate><topic>Analysis</topic><topic>Enzymes</topic><topic>Escherichia coli</topic><topic>Ethylenediaminetetraacetic acid</topic><topic>Lactase</topic><topic>Lactose</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Xin, Yongping</creatorcontrib><creatorcontrib>Guo, Tingting</creatorcontrib><creatorcontrib>Zhang, Yi</creatorcontrib><creatorcontrib>Wu, Jiapeng</creatorcontrib><creatorcontrib>Kong, Jian</creatorcontrib><collection>Gale In Context: Science</collection><jtitle>Applied microbiology and biotechnology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Xin, Yongping</au><au>Guo, Tingting</au><au>Zhang, Yi</au><au>Wu, Jiapeng</au><au>Kong, Jian</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A new [beta]-galactosidase extracted from the infant feces with high hydrolytic and transgalactosylation activity</atitle><jtitle>Applied microbiology and biotechnology</jtitle><date>2019-10-01</date><risdate>2019</risdate><volume>103</volume><issue>20</issue><spage>8439</spage><pages>8439-</pages><issn>0175-7598</issn><eissn>1432-0614</eissn><abstract>A [beta]-galactosidase ([beta]-Gal.sub.INF) was directly isolated from feces of an 8-month-old infant. Mass spectrum analysis showed [beta]-Gal.sub.INF with coverage over 50% to the [beta]-galactosidase from Bifidobacterium longum EK3. Accordingly, the [beta]-gal.sub.INF was amplified from the feces metagenomic DNA by degenerate primers. After overexpressed in Escherichia coli, the [beta]-Gal.sub.INF was purified and biochemical characterized. [beta]-Gal.sub.INF existed as homotetramer and homodimer, whose activity (optimal at 50 °C, pH 6.5) was exhilaratingly increased to 484% by artificial intestinal juice. The K.sub.m and V.sub.max values for oNPG and lactose were 20.95 ± 2.76 mM, 5004.50 ± 318.8 [mu]mol min.sup.-1 mg.sup.-1 and 140.2 ± 17.7 mM, 293.1 ± 14.7 [mu]mol min.sup.-1 mg.sup.-1, respectively. The production rate of galacto-oligosaccharides by [beta]-Gal.sub.INF from 20% lactose at 50 °C was 33.4 ± 0.67%. These results suggested the [beta]-Gal.sub.INF with high hydrolytic and transgalactosylation activity from the infant intestinal has great potential as infant lactase preparation. Moreover, this study provided a new way for exploring undetected enzymes by uncultured-dependent methods.</abstract><pub>Springer</pub><doi>10.1007/s00253-019-10092-x</doi><tpages>10</tpages></addata></record> |
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| ispartof | Applied microbiology and biotechnology, 2019-10, Vol.103 (20), p.8439 |
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| source | Springer Nature - Complete Springer Journals |
| subjects | Analysis Enzymes Escherichia coli Ethylenediaminetetraacetic acid Lactase Lactose |
| title | A new [beta]-galactosidase extracted from the infant feces with high hydrolytic and transgalactosylation activity |
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