Expression of Recombinant Galactose Oxidase by Pichia pastoris

2000 ◽  
Vol 20 (1) ◽  
pp. 105-111 ◽  
Author(s):  
Mei M. Whittaker ◽  
James W. Whittaker
Keyword(s):  
Pichia Pastoris ◽  
Galactose Oxidase

scholarly journals Effects of temperature and glycerol and methanol‐feeding profiles on the production of recombinant galactose oxidase in Pichia pastoris

2014 ◽  
Vol 30 (3) ◽  
pp. 728-735 ◽  
Author(s):  
George E. Anasontzis ◽  
Margarita Salazar Penã ◽  
Oliver Spadiut ◽  
Harry Brumer ◽  
Lisbeth Olsson
Keyword(s):  
Pichia Pastoris ◽  
Galactose Oxidase ◽  
Methanol Feeding ◽  
Effects Of Temperature

Functional Fractionation of Platelets: Aggregation Kinetics and Glycoprotein Labeling of Differing Platelet Populations

1982 ◽  
Vol 48 (02) ◽  
pp. 211-216 ◽  
Author(s):  
V M Haver ◽  
A R L Gear
Keyword(s):  
Galactose Oxidase ◽  
Aggregation Kinetics ◽  
Maximal Rate ◽  
Membrane Glycoproteins ◽  
Whole Cells ◽  
Reversible Aggregation ◽  
Significant Difference ◽  
Small Doses ◽  
Major Loss ◽  
Kinetics Of

SummaryPlatelet heterogeneity has been studied with a technique called functional fractionation which employs gentle centrifugation to yield subpopulations (“reactive” and “less-reactive” platelets) after exposure to small doses of aggregating agent. Aggregation kinetics of the different platelet populations were investigated by quenched-flow aggregometry. The large, “reactive” platelets were more sensitive to ADP (Ka = 1.74 μM) than the smaller “less-reactive” platelets (Ka = 4.08 μM). However, their maximal rate of aggregation (Vmax, % of platelets aggregating per sec) of 23.3 was significantly lower than the “less-reactive” platelets (Vmax = 34.7). The “reactive” platelets had a 2.2 fold higher level of cyclic AMP.Platelet glycoproteins were labeled using the neuraminidase-galactose oxidase – [H3]-NaBH4 technique. When platelets were labeled after reversible aggregation, the “reactive” platelets showed a two-fold decrease in labeling efficiency (versus control platelets). However, examination of whole cells or membrane preparations from reversibly aggregated platelets revealed no significant difference in Coomassie or PAS (Schiff) staining.These results suggest that the large, “reactive” platelets are more sensitive to ADP but are not hyperaggregable in a kinetic sense. Reversible aggregation may cause a re-orientation of membrane glycoproteins that is apparently not characterized by a major loss of glycoprotein material.

Optimization of fermentation conditions for the production of recombinant hpdgf-bb in Pichia pastoris

2015 ◽  
Vol 37 (1se) ◽  
Author(s):  
Duong Long Duy ◽  
Pham Minh Vu ◽  
Nguyen Tri Nhan ◽  
Tran Linh Thuoc ◽  
Dang Thi Phuong Thao
Keyword(s):  
Pichia Pastoris ◽  
Fermentation Conditions ◽  
Optimization Of Fermentation

Cloning and Expression of Recombinant Human Insulin Gene in Pichia pastoris

2019 ◽  
Vol 10 (01) ◽  
Author(s):  
Rafid A. Abdulkareem
Keyword(s):  
Pichia Pastoris ◽  
Human Insulin ◽  
Expression Vector ◽  
Expression System ◽  
Insulin Gene ◽  
Cloning And Expression ◽  
Pcr Analysis ◽  
Major Band ◽  
Human Insulin Gene ◽  
Pichia Pastoris Expression System

The main goal of the current study was cloning and expression of the human insulin gene in Pichia pastoris expression system, using genetic engineering techniques and its treatment application. Total RNA was purified from fresh normal human pancreatic tissue. RNA of good quality was chosen to obtain a first single strand cDNA. Human preproinsulin gene was amplified from cDNA strand, by using two sets of specific primers contain EcoR1 and Notl restriction sites. The amplified preproinsulin gene fragment was double digested with EcoRI and Not 1 restriction enzymes, then inserted into pPIC9K expression vector. The new pPIC9K-hpi constructive expression vector was transformed by the heat-shock method into the E.coli DH5α competent cells. pPic9k –hpi, which was propagated in the positive transformant E. coli cells, was isolated from cells and then linearised by restriction enzyme SalI, then transformed into Pichia pastoris GS115 using electroporation method. Genomic DNA of His+ transformants cell was extracted and used as a template for PCR analysis. The results showed, that the pPic9k – hpi was successfully integrated into the P. pastoris genome, for selected His+ transformants clones on the anticipated band at 330 bp, which is corresponded to the theoretical molecular size of the human insulin gene. To follow the insulin expression in transformans, Tricine–SDS gel electrophoresis and Western blot analysis were conducted. The results showed a successful expression of recombinant protein was detected by the presence of a single major band with about (5.8 KDa) on the gel. These bands correspond well with the size of human insulin with the theoretical molecular weight (5.8 KDa).

SECRETED EXPRESSION OF LITOPENAEUS VANNAMEI LYSOZYME GENE IN PICHIA PASTORIS AND ITS BIOACTIVITY DETECTION

2010 ◽  
Vol 36 (6) ◽  
pp. 1091-1096
Author(s):  
Shu-Guang BIAN ◽  
Hua-Xin CHEN ◽  
Peng JIANG ◽  
Hai-Bo ZHANG ◽  
Zhao-Pu LIU ◽  
...  
Keyword(s):  
Pichia Pastoris ◽  
Litopenaeus Vannamei ◽  
Lysozyme Gene ◽  
Secreted Expression

Expression and Characteristics of Phytases from Obesumbacterium proteus in Pichia pastoris Yeast

2018 ◽  
Vol 34 (4) ◽  
pp. 18-25 ◽  
Author(s):  
T.L. Gordeeva ◽  
◽  
L.N. Borshchevskaya ◽  
A.N. Kalinina ◽  
S.P. Sineoky ◽  
...  
Keyword(s):  
Pichia Pastoris ◽  
Pichia Pastoris Yeast
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