« Previous Abstract"The dynamics of cereal cyst nematode infection differ between susceptible and resistant barley cultivars and lead to changes in (1,3;1,4)-beta-glucan levels and HvCslF gene transcript abundance"    Next AbstractAdult House Fly (Diptera: Muscidae) Response to Black Soldier Fly (Diptera: Stratiomyidae) Associated Substrates and Potential Volatile Organic Compounds Identification »

Int J Biol Macromol

Title:		High level production of active streptokinase in Pichia pastoris fed-batch culture
Author(s):		Adivitiya; Dagar VK; Devi N; Khasa YP;
Address:		"Department of Microbiology, University of Delhi South Campus, New Delhi 110 021, India. Department of Microbiology, University of Delhi South Campus, New Delhi 110 021, India. Electronic address: yogi110@gmail.com"
Journal Title:		Int J Biol Macromol
Year:		2016
Volume:		20151126
Issue:
Page Number:		50 - 60
DOI:		10.1016/j.ijbiomac.2015.11.062
ISSN/ISBN:		1879-0003 (Electronic) 0141-8130 (Linking)
Abstract:		"Streptokinase is a biological macromolecule involved in dissolution of fibrin blood clot and favourably used in various clinical applications. This protein is poorly expressed in soluble form due to its toxic effects on host physiology. The extracellular expression of recombinant streptokinase (SK) with and without 6xHis tag was obtained by cloning its gene under the alpha-mating factor signal sequence and alcohol inducible AOX1 promoter. Host-vector combinations were optimized to select a hyper producer. From shake flask optimization studies, a maximum expression of 582 mg/L of rSK (non-tagged) and 538 mg/L of rSK-His (His-tagged) protein was obtained when cells were induced at OD600 of 20. The high cell density fermentation increased the volumetric product concentration of rSK-His to a level of 4.25 g/L with a 7.9 folds increase from shake flask results. The specific product yield (YP/X) was 49.75 mg/g DCW along with a high volumetric productivity of 57.43 mg/L/h. The protein was predicted to have 15.43% alpha-helix and 26.43% beta-sheet with tryptophan emission maxima of around 347 nm. The highest specific activity of rSK-His was 64,903 IU/mg with 1.48 folds purification whereas specific activity of rSK was 55,240 IU/mg with 1.22 folds purification"
Keywords:		"Batch Cell Culture Techniques/methods Cloning, Molecular/methods Fermentation/genetics Gene Expression/genetics Mating Factor Peptides/genetics/metabolism Pichia/*genetics/*metabolism Promoter Regions, Genetic/genetics Protein Sorting Signals/genetics Rec;"
Notes:		"MedlineAdivitiya Dagar, Vikas Kumar Devi, Nirmala Khasa, Yogender Pal eng Research Support, Non-U.S. Gov't Netherlands 2015/12/04 Int J Biol Macromol. 2016 Feb; 83:50-60. doi: 10.1016/j.ijbiomac.2015.11.062. Epub 2015 Nov 26"