Investigation of Bacterial Cellulose Biosynthesis Mechanism in Gluconoacetobacter hansenii.

ISRN Microbiology Pub Date : 2014-03-16 eCollection Date: 2014-01-01 DOI:10.1155/2014/836083
Bhavna V Mohite, Satish V Patil
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Abstract

The present study explores the mechanism of cellulose biosynthesis in Gluconoacetobacter hansenii. The cellulose synthase enzyme was purified as membrane fraction and solubilized by treatment with 0.1% digitonin. The enzyme was separated by native-gel electrophoresis and β -D-glucan analysis was carried out using in vitro gel assay. The cellulose synthase has glycoprotein nature and composed two polypeptide subunits of 93 KDa and 85 KDa. The confirmation of β -1,4-glucan (cellulose) was performed in whole and hydrolyzed monomeric sugar form. Tinopal and Congo red were used for cellulose detection on the gel. Thus the in vitro cellulose synthesis assay with cell free enzyme fraction was attempted to improve the understanding of cellulose biosynthesis.

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Gluconoacetobacter hansenii 细菌纤维素生物合成机制研究
本研究探讨了汉森葡萄糖乙酰杆菌纤维素生物合成的机制。纤维素合成酶以膜部分纯化,并用 0.1% 的地高辛处理增溶。酶通过原生凝胶电泳进行分离,并使用体外凝胶测定法对β-D-葡聚糖进行分析。纤维素合成酶具有糖蛋白性质,由 93 KDa 和 85 KDa 的两个多肽亚基组成。β-1,4-葡聚糖(纤维素)以整体和水解单糖形式进行确认。凝胶上的纤维素检测使用了锡醛和刚果红。因此,尝试用细胞游离酶部分进行体外纤维素合成测定,以增进对纤维素生物合成的了解。
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