首页|生物信息学分析葡聚糖蔗糖酶的结构和功能特性

生物信息学分析葡聚糖蔗糖酶的结构和功能特性

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葡聚糖蔗糖酶是一种重要的糖基转移酶(Glucosyltransferase,GTF),对于乳酸菌合成胞外多糖具有关键作用,能够以蔗糖为底物合成葡聚糖或低聚糖,是乳酸菌(Lactic acid bacteria,LAB)合成胞外多糖(Exopolysaccharide,EPS)的关键酶蛋白.然而乳酸菌代谢系统复杂,EPS生物合成机制未得到全面解析,限制了 EPS的应用.为了进一步解析乳酸菌EPS的生物合成机制,表征不同来源葡聚糖蔗糖酶的结构,探明酶的催化调控机制是必要手段.本研究以肠膜明串珠菌(Leuconostoc mesenteroides)DRP105的葡聚糖蔗糖酶gtfB基因为目标序列,利用生物信息学技术对葡聚糖蔗糖酶GtfB结构和功能进行了预测分析.研究结果显示:GtfB属于GH70家族,是一种胞外酶,不存在跨膜区,含有7个保守区域和39条重复序列.其理论分子量为308986.21,理论等电点为4.59,属于酸性蛋白质.磷酸化位点含110个Thr、89个Ser、53个Tyr,GtfB含有5个结构域,呈U型折叠,活性中心在A结构域,含有(β/α)8桶状结构.分子对接预测分析表明,蔗糖与GtfB的活性口袋结合紧密.这些结果初步探明了 GtfB具有葡聚糖蔗糖酶的特征结构和功能,证明其在乳酸菌EPS合成途径中的关键调控作用,为其在EPS生产过程中的应用提供了理论基础.
Analysis of Structure and Functional Properties of Glucansucrase Based on Bioinformatics Method
Glucansucrase is a glycosyltransferase(GTF)that can synthesize dextran or oligosaccharides with sucrose as a substrate,and is a key enzyme protein for the synthesis of exopolysaccharide(EPS)by lactic acid bacteria(LAB).However,the metabolic system of LAB is complex,and the mechanism of EPS biosynthesis has not been elucidated,which limits the development of EPS.In order to further analyze the biosynthetic mechanism of LAB EPS and characterize the structure of dextransucrase from different sources,it is necessary to explore the catalytic regulation mechanism of enzyme.In this study,the structure and function of glucansucrase GtfB from Leuconostoc mesenteroides DRP105 were predicted with bioinformatics techniques.The results showed that GtfB belonged to the GH70 family,which was an extracellular enzyme with no transmembrane region,containing 7 conserved regions and 39 repeats.The theoretical molecular weight of GtfB was 308986.21,and the theoretical isoelectric point was 4.59,which was an acidic protein.The phosphorylation site contained 110 Thr,89 Ser,and 53 Tyr,and GtfB contained 5 domains,which were U-folded,and the active center was in the A domain,containing(β/α)8 barrel-like structures.Molecular docking prediction analysis showed that sucrose was tightly bound to the active pocket of GtfB.The results of this study preliminarily revealed the characteristic structure and function of GtfB as a glucanase,proving that this enzyme is a key regulatory enzyme in the EPS synthesis pathway of LAB and has potential application value in the EPS production process.

Leuconostoc mesenteroidesglucansucrasebioinformatics analysisstructure predictionmolecular docking

蔡华扬、于连升、李腾鑫、林宜萌、杜仁鹏

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黑龙江大学,生命科学学院,农业微生物技术教育部工程研究中心,黑龙江省寒区植物基因与生物发酵重点实验室,黑龙江省普通高校微生物重点实验室,哈尔滨 150080

河北环境工程学院,河北省农业生态安全重点实验室,河北秦皇岛 066102

肠膜明串珠菌 葡聚糖蔗糖酶 生物信息学分析 结构预测 分子对接

黑龙江省自然科学基金优秀青年基金中国博士后科学基金黑龙江省博士后资助项目新时代龙江优秀硕士、博士学位论文项目河北省农业生态安全重点实验室开放基金黑龙江省省属高校基本科研业务费专项

YQ2021C0302022MD713755LBH-Z21082LJYXL2022-0202023SYSJJ172022-KYYWF-1075

2024

中国农学通报
中国农学会

中国农学通报

CSTPCD
影响因子：0.891
ISSN：1000-6850
年,卷(期)：2024.40(18)