重组人α-葡萄糖苷酶在HEK293细胞中的表达

IF 6.2 1区农林科学 Q1 AGRICULTURE, MULTIDISCIPLINARY

Journal of Agricultural and Food Chemistry Pub Date : 2024-12-23 DOI:10.1021/acs.jafc.4c06902

So Nishimoto, Anaïs Debarbat, Yuki Ikeda, Emi Arikawa, Yuki Odagaki, Haruna Yano, Ying Qiao, Masaaki Ito, Toshiyuki Kimura, Teisuke Takita, Kiyoshi Yasukawa

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引用次数: 0

摘要

在哺乳动物中，肠道α-葡萄糖苷酶以麦芽糖酶-葡萄糖淀粉酶复合物（MGAM）和蔗糖-异麦芽糖酶复合物（SI）的形式存在。在这项研究中，我们在人胚胎肾293 （HEK293）细胞中短暂表达了人MGAM和SI。在pH 6.0和37°C下，表达mgam的HEK293细胞提取物（MGE）表现出麦芽糖酶、葡萄糖淀粉酶和异麦芽糖酶的活性，但不表现出蔗糖酶的活性，而表达si的HEK293细胞提取物（SIE）表现出蔗糖酶、异麦芽糖酶和麦芽糖酶的活性，但不表现出葡萄糖淀粉酶的活性。麦芽糖水解MGE的表观Km值是麦芽糖、蔗糖和异麦芽糖水解SIE的14-26%。蔗糖和异麦芽糖水解MGE和SIE的表观Vmax分别为麦芽糖水解的0%和6%和10%和42%。这些结果表明，MGAM和SI的麦芽糖酶活性高于蔗糖酶和异麦芽糖酶。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

Expression of Recombinant Human α-Glucosidase in HEK293 Cells

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Expression of Recombinant Human α-Glucosidase in HEK293 Cells

In mammals, intestinal α-glucosidase exists as a maltase–glucoamylase complex (MGAM) and a sucrase–isomaltase complex (SI). In this study, we transiently expressed human MGAM and SI in human embryonic kidney 293 (HEK293) cells. At pH 6.0 and 37 °C, the MGAM-expressing HEK293 cells extract (MGE) exhibited maltase, glucoamylase, and isomaltase activities but not sucrase activity, whereas the SI-expressing HEK293 cells extract (SIE) exhibited sucrase, isomaltase, and maltase activities but not glucoamylase activity. The apparent K_m value of the MGE for maltose hydrolysis was 14–26% of that of the SIE for maltose, sucrose, and isomaltose hydrolysis. The respective apparent V_max values of the MGE and SIE for sucrose and isomaltose hydrolysis were 0% and 6% and 10% and 42% of those for maltose hydrolysis. These results indicated that the maltase activities of MGAM and SI were higher than those of sucrase and isomaltase.

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来源期刊

Journal of Agricultural and Food Chemistry 农林科学-农业综合

CiteScore

9.90

自引率

8.20%

发文量

1375

审稿时长

2.3 months

期刊介绍： The Journal of Agricultural and Food Chemistry publishes high-quality, cutting edge original research representing complete studies and research advances dealing with the chemistry and biochemistry of agriculture and food. The Journal also encourages papers with chemistry and/or biochemistry as a major component combined with biological/sensory/nutritional/toxicological evaluation related to agriculture and/or food.