Method of reducing glycosylation of proteins, processes and proteins thereof

US8778659B2 · kind B2 · utility

Assignee

• Biocon Ltd. · IN

Inventors

• Nagaraj Govindappa · Bengaluru, IN
• Komal N. Kanojia · Bengaluru, IN
• Krishnamurthy Venkatesan · Viralimalai, IN
• Nitesh Dave · Bengaluru, IN
• Mukesh Babuappa Patale · Jamkhed, IN
• Sanjay Tiwari · Meridian, US
• Kedarnath Sastry · Savanuru, IN
• Harish Iyer · Bengaluru, IN

Key dates

• Filing date: Mar 29, 2010
• Grant date: Jul 15, 2014
• Expiry date: Mar 29, 2030

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12N15/815
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

The disclosure relates to method of reducing O-glycosylation levels of the insulin or insulin analog precursor molecule produced by Pichia sp. The present disclosure provides genetically engineered knock-out strains of methylotrophic yeast including Pichia and especially Pichia pastoris by disruption of Protein mannosyl transferase (PMT) genes and rendering them capable of producing heterologous proteins with reduced glycosylation. Vectors, which comprise coding sequences for PMT1, PMT2, PMT4, PMT5, and PMT6 genes, for transforming methylotrophic yeasts are contemplated by the present disclosure. PMT inactivated strains of this disclosure have been deposited at MTCC, Chandigarh. The strains are PMT1/GS115 (MTCC 5515), PMT4/GS115 (MTCC 5516), PMT5/GS115 (MTCC 5517) and PMT6/GS115 (MTCC 5518).