Biocon Limited

Krishnamurthy Venkatesan

Sanjay Tiwari

Harish Iyer

Mukesh Babuappa Patale

Nitesh Dave

Kedarnath Nanjund Sastry

Nagaraj Govindappa

Anuj Goel

Partha Hazra

Komal N Kanojia

Nita Roy

Goldy Sachdev

Vivekanandan Kannan

Anoop Vasudevan

Anupama Jagadish

Komal Kanojia

The invention relates to methods of separation and/or purification of impurities yielding a purified heterologous protein product devoid of related impurities or with substantially minimal quantities of such glycosylated impurities. More specifically, the invention relates to the identification of glycosylated forms of insulin analogues such as glargine impurities characterized post expression in yeast based systems such as. The invention also relates to methods used to clone gene encoding the protein insulin glargine; inserting the related gene in a suitable yeast host; producing culture of the recombinant strain, stimulating expression of the heterologous polypeptide, its secretion and purification post fermentation and related enzymatic conversions.

Method of obtaining a purified, biologically active heterologous protein

The disclosure relates to method of reducing O-glycosylation levels of the insulin or insulin analog precursor molecule produced bysp. The present disclosure provides genetically engineered knock-out strains of methylotrophic yeast includingand especiallyby 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).

Method of reducing glycosylation of proteins, processes and proteins thereof

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Tamil Nadu, India

Krishnamurthy Venkatesan