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首页> 外文期刊>Protein Expression and Purification >Cloning and characterization of a gene encoding trehalose phosphorylase (TP) from Pleurotus sajor-caju
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Cloning and characterization of a gene encoding trehalose phosphorylase (TP) from Pleurotus sajor-caju

机译：侧耳侧耳海藻糖磷酸化酶（TP）基因的克隆与鉴定

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摘要

Complementary DNA for a gene encoding trehalose phosphorylase (TP) that reversibly catalyzes trehalose synthesis and degradation from alpha-glucose-1-phosphate (alpha-Glc-1-P) and glucose was cloned from Pleurotus sajor-caju. The cDNA of P. sajor-caju TP (designated PsTP, GenBank Accession No. AF149777) encodes a polypeptide of 751 amino acids with a deduced molecular mass of 83.7 kDa. The PsTP gene is expressed in mycelia, pilei, and stipes of fruiting bodies. Trehalose phosphorylase PsTP was purified from PsTP-transformed Escherichia coli. The enzyme catalyzes both the phosphorolysis of trehalose to produce alpha-Glc-1-P and glucose, and the synthesis of trehalose. The apparent K-m values for trehalose and Pi in phosphorolytic reaction at pH 7.0 were 74.8 and 5.4mM, respectively. The PsTP gene complemented Saccharomyces cerevisiae Deltatps1, Deltatps2 double-mutant cells, allowing their growth on glucose medium. Furthermore, yeast transformed with PsTP produced 2-2.5-fold more trehalose than non-transformants or cells transformed with empty vector only. (C) 2003 Elsevier Science (USA). All rights reserved. [References: 33]
机译：从平菇中克隆了编码海藻糖磷酸化酶（TP）的基因的互补DNA，该酶可逆性催化海藻糖的合成和降解，由α-葡萄糖-1-磷酸（α-Glc-1-P）和葡萄糖组成。 Sajor-caju TP的cDNA（指定为PsTP，GenBank登录号AF149777）编码751个氨基酸的多肽，推导分子量为83.7 kDa。 PsTP基因在子实体的菌丝体，桩和柄中表达。海藻糖磷酸化酶PsTP从PsTP转化的大肠杆菌中纯化。该酶既催化海藻糖的磷酸分解以产生α-Glc-1-P和葡萄糖，又催化海藻糖的合成。在pH 7.0的磷解反应中，海藻糖和Pi的表观K-m值分别为74.8和5.4mM。 PsTP基因补充了酿酒酵母Deltatps1，Deltatps2双突变细胞，使其在葡萄糖培养基上生长。此外，用PsTP转化的酵母产生的海藻糖比非转化子或仅用空载体转化的细胞多2-2.5倍。 （C）2003 Elsevier Science（美国）。版权所有。 [参考：33]

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