Characterization of a novel type of glycogen-degrading amylopullulanase from Lactobacillus crispatus
Characterization of a novel type of glycogen-degrading amylopullulanase from Lactobacillus crispatus
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卷曲乳杆菌中新型糖原降解淀粉支链淀粉酶的表征
DOI：
10.1007/s00253-022-11975-2
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发表时间：
2022-05
影响因子：
5
通讯作者：
Jin Zhong
中科院分区：
文献类型：
--
作者：
Jie Zhang;Lili Li;Tong Zhang;Jin Zhong
糖原是人类阴道微生物群利用的主要碳水化合物之一，其通常由乳杆菌属（Lactobacillus）尤其是乳酸杆菌（L.卷曲的计算机模拟分析预测I型普鲁兰酶参与L.卷曲的普鲁兰酶的生物化学和遗传特性仍然需要确定。在此，我们重新鉴定了L.通过生物化学测定，GlgU的最适pH为4.0 ~ 4.5，能水解糖原生成低分子量的麦芽低聚糖。实际上，GlgU是一种II型普鲁兰酶（淀粉普鲁兰酶），只有一个催化结构域，对α-1，4和α-1，6-糖苷键都具有底物特异性。在系统发育上，GlgU与乳酸杆菌中已知的淀粉酶和支链淀粉酶（包括淀粉支链淀粉酶）明显不同。此外，我们证实了glgU在非糖原利用乳酸杆菌菌株中的催化活性，证明了glgU在糖原代谢中的重要作用。总的来说，这项研究表征了一种新型的淀粉普鲁兰酶，有助于了解人类阴道微生物群优势物种的糖原利用机制。关键点：· GlgU是一种II型普鲁兰酶，而不是以前预测的I型普鲁兰酶。· GlgU能够将糖原完全水解成麦芽寡糖。· GlgU在细胞外糖原代谢中起关键作用。
Glycogen is one of the major carbohydrates utilized by the human vaginal microbiota, which is commonly dominated by Lactobacillus, especially L. crispatus. An in silico analysis predicted that a type I pullulanase was involved in glycogen degradation in L. crispatus. The biochemical and genetic properties of the pullulanase still need to be determined. Here, we de novo identified the glycogen (Glg)-utilization enzyme (named GlgU) from L. crispatus through a biochemical assay. GlgU was optimally active at acidic pH, approximately 4.0 ~ 4.5, and was able to hydrolyze glycogen into low-molecular-weight malto-oligosaccharides. Actually, GlgU was a type II pullulanase (amylopullulanase) with just one catalytic domain that possessed substrate specificity toward both α-1,4 and α-1,6-glucosidic bonds. Phylogenetically, GlgU was obviously divergent from the known amylases and pullulanases (including amylopullulanases) in lactobacilli. In addition, we confirmed the catalytic activity of glgU in a nonglycogen-utilizing lactobacilli strain, demonstrating the essential role of glgU in glycogen metabolism. Overall, this study characterized a novel type of amylopullulanases, contributing to the knowledge of the glycogen utilization mechanism of the dominant species of human vaginal microbiota. KEY POINTS: • GlgU was a type II pullulanase, not a type I pullulanase predicted before. • GlgU was able to completely hydrolyze glycogen into malto-oligosaccharides. • GlgU played a key role in the metabolism of extracellular glycogen.
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影响因子：
15.5
作者：
C. van der Veer;R. Hertzberger;S. Bruisten;Hanne L. P. Tytgat;Jorne Swanenburg;Alie de Kat Angelino-Bart;F. Schuren;D. Molenaar;G. Reid;H. D. de Vries;R. Kort
通讯作者：
C. van der Veer;R. Hertzberger;S. Bruisten;Hanne L. P. Tytgat;Jorne Swanenburg;Alie de Kat Angelino-Bart;F. Schuren;D. Molenaar;G. Reid;H. D. de Vries;R. Kort
DOI：
10.1016/j.ijbiomac.2018.01.166
发表时间：
2018-06
影响因子：
8.2
作者：
R. Tester;F. Al-Ghazzewi
通讯作者：
R. Tester;F. Al-Ghazzewi
DOI：
10.3109/10425170109042048
发表时间：
2001-01
期刊：
DNA Sequence
影响因子：
--
作者：
J. Morlon‐Guyot;F. Mucciolo-Roux;R. R. Sanoja-R.;J. Guyot
通讯作者：
J. Morlon‐Guyot;F. Mucciolo-Roux;R. R. Sanoja-R.;J. Guyot
超越能量储存：糖原代谢在健康和疾病中的作用。
影响因子：
5.4
作者：
Zhang Huafeng;Ma Jingwei;Tang Ke;Huang Bo
通讯作者：
Huang Bo
DOI：
10.1515/amylase-2021-0001
发表时间：
2021-01
期刊：
Amylase
影响因子：
--
作者：
Carolina Rodríguez-Saavedra;R. Rodríguez-Sanoja;D. Guillén;C. Wacher;G. Díaz-Ruiz
通讯作者：
Carolina Rodríguez-Saavedra;R. Rodríguez-Sanoja;D. Guillén;C. Wacher;G. Díaz-Ruiz