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A Novel Gene Encoding Xanthan Lyase of Paenibacillus alginolyticus Strain XL-1

机译:编码解藻杆状杆菌XL-1的黄原酸裂解酶的新基因。

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摘要

Xanthan-modifying enzymes are powerful tools in studying structure-function relationships of this polysaccharide. One of these modifying enzymes is xanthan lyase, which removes the terminal side chain residue of xanthan. In this paper, the cloning and sequencing of the first xanthan lyase-encoding gene is described, i.e., the xalA gene, encoding pyruvated mannose-specific xanthan lyase of Paenibacillus alginolyticus XL-1. The xalA gene encoded a 100,823-Da protein, including a 36-amino-acid signal sequence. The 96,887-Da mature enzyme could be expressed functionally in Escherichia coli. Like the native enzyme, the recombinant enzyme showed no activity on depyruvated xanthan. Compared to production by P. alginolyticus, a 30-fold increase in volumetric productivity of soluble xanthan lyase was achieved by heterologous production in E. coli. The recombinant xanthan lyase was used to produce modified xanthan, which showed a dramatic loss of the capacity to form gels with locust bean gum.
机译:黄原胶修饰酶是研究这种多糖的结构-功能关系的有力工具。这些修饰酶之一是黄原胶裂解酶,其去除了黄原胶的末端侧链残基。在本文中,描述了第一个黄原酸裂解酶编码基因的克隆和测序,即xalA基因,它编码解藻性杆状芽胞杆菌XL-1的丙酮酸甘露糖特异性黄原酸裂解酶。 xalA基因编码100,823-Da蛋白,包括36个氨基酸的信号序列。 96,887-Da成熟酶可以在大肠杆菌中功能性表达。与天然酶一样,重组酶对去丙酮化的黄原没有活性。与解藻毕赤酵母的生产相比,通过在大肠杆菌中进行异源生产,可溶性黄原酸裂解酶的体积生产率提高了30倍。重组黄原胶裂解酶用于生产修饰的黄原胶,其显示出与刺槐豆胶形成凝胶的能力显着降低。

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