Asparagine as a nitrogen source for improving the secretion of mouse ?-amylase inSaccharomyces cerevisiae protease A-deficient strains

Autor: Bi-Dar Wang, Tsong-Teh Kuo, Dz-chi Chen, Pei-Yin Chou
Rok vydání: 2000
Předmět:
Zdroj: Yeast. 16:207-217
ISSN: 1097-0061
0749-503X
DOI: 10.1002/(sici)1097-0061(200002)16:3<207::aid-yea518>3.0.co;2-c
Popis: A modified chemically defined medium was achieved by using asparagine as a nitrogen source to increase the production of secreted mouse α-amylase in several Saccharomyces cerevisiae protease A-deficient (pep4) strains. The specific productivity (quantity) and the 53 kDa non-glycosylated active form (quality) of mouse salivary α-amylase in liquid medium containing asparagine was remarkably improved compared to media containing other nitrogen sources, including ammonium sulphate, glutamic acid, arginine, casamino acids, yeast extract and peptone. Similar improvement was also observed on starch solid agar regarding the clarity and size of the halo zone formed by α-amylase activity. Compared with ammonium sulphate, advantages of using asparagine as the nitrogen source in liquid or solid medium included increasing the cell mass of test strains, recovering the viability of protease-deficient strains to levels similar to the wild-type strain, and increasing the copy number of the mouse α-amylase expression vector in test strains. In turn, these advantages apparently contributed to the increase of secretion of mouse α-amylase in several test strains and especially in the protease A-deficient strains. In addition to demonstrating the use of modified chemically defined medium to improve the quality and quantity of secreted mouse α-amylase, this study also provides a new strategy to improve the secretion of heterologous proteins in protease A deficient strains. Copyright © 2000 John Wiley & Sons, Ltd.
Databáze: OpenAIRE
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