Optimization of Production Conditions of His-taqqed pro-Microbial Transglutaminase Enzyme in Pichia pastoris

Abstract

Transglutaminases are enzymes involved in various biological processes and found in many organisms in nature. They form a cross-link by catalyzing the formation of covalent bonds between the amino acids glutamine and lysine. Due to these properties, they are used to improve the physical and chemical properties of food products. In this study, the production conditions of microbial transglutaminase (MTGase) enzyme, used in the food industry, in Pichia pastoris (P. pastoris) yeast under the structural GAP promoter were optimized. For this purpose, the production of the pro-MTGase enzyme under nine different conditions consisting of three different temperatures (20°C, 25°C, 28°C) and three different pHs (pH 3, 5.75, 7.44) was carried out by shake flask experiments. The results obtained after 40 hours of protein production showed that; the optimum conditions were 25°C temperature and pH 7.44 for pro-MTGase production.

Keywords

• microbial transglutaminase,recombinant protein production,Pichia pastoris

His-etiketli pro-Mikrobiyal Transglutaminaz Enziminin Pichia pastoris’te Üretim Şartlarının Optimizasyonu

Abstract

Transglutaminazlar doğada birçok organizmada bulunan ve çeşitli biyolojik işlemlerde rol alan enzimlerdir. Glutamin ve lizin aminoasitleri arasında kovalent bağ oluşumunu katalizleyerek çapraz bağ oluştururlar. Bu özelliklerinden dolayı gıda ürünlerinin fiziksel ve kimyasal özelliklerini geliştirmek amacıyla kullanılırlar. Bu çalışmada, gıda endüstrisinde kullanılan mikrobiyal transglutaminaz (MTGaz) enziminin, Pichia pastoris (P. pastoris) mayasında yapısal GAP promotoru altında üretim şartlarının optimizasyonu yapılmıştır. Bu amaçla, üç farklı sıcaklık (20°C, 25°C, 28°C) ve üç farklı pH’dan (pH 3, 5.75, 7.44) oluşan dokuz farklı koşul altında pro-MTGaz enziminin üretimi çalkalamalı erlenmayerlerde yapılmıştır. 40 saatlik protein üretimi sonrasında elde edilen sonuçlar, pro-MTGaz üretimi için 25°C sıcaklık ve 7.44 pH’nın optimum koşullar olduğunu göstermiştir.

Keywords

• mikrobiyal transglutaminaz,rekombinant protein üretimi,Pichia pastoris

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