scholarly journals Cloning and expression of a family 10 xylanase gene (Aoxyn10) from Aspergillus oryzae in Pichia pastoris

2013 ◽  
Vol 59 (6) ◽  
pp. 405-415 ◽  
Author(s):  
Xin Yin ◽  
Yan-Yan Gong ◽  
Jun-Qing Wang ◽  
Cun-Duo Tang ◽  
Min-Chen Wu
Keyword(s):  
Pichia Pastoris ◽  
Aspergillus Oryzae ◽  
Cloning And Expression ◽  
Xylanase Gene ◽  
Family 10 Xylanase

Cloning and Expression of a Novel Xylanase Gene (Auxyn11D) from Aspergillus usamii E001 in Pichia pastoris

2012 ◽  
Vol 167 (8) ◽  
pp. 2198-2211 ◽  
Author(s):  
Huimin Zhang ◽  
Minchen Wu ◽  
Jianfang Li ◽  
Shujuan Gao ◽  
Yanjun Yang
Keyword(s):  
Pichia Pastoris ◽  
Cloning And Expression ◽  
Xylanase Gene ◽  
Aspergillus Usamii

Cloning and expression of GH11 xylanase gene from Aspergillus fumigatus MKU1 in Pichia pastoris

2009 ◽  
Vol 108 (1) ◽  
pp. 24-29 ◽  
Author(s):  
Marimuthu Jeya ◽  
Sairam Thiagarajan ◽  
Jung-Kul Lee ◽  
Paramasamy Gunasekaran
Keyword(s):  
Pichia Pastoris ◽  
Aspergillus Fumigatus ◽  
Cloning And Expression ◽  
Xylanase Gene

Cloning and expression of the Aspergillus oryzae glucan 1,3-beta-glucosidase A (exgA) in Pichia pastoris

2012 ◽  
Vol 34 (10) ◽  
pp. 1937-1943 ◽  
Author(s):  
Nassapat Boonvitthya ◽  
Phatrapan Tanapong ◽  
Patcharaporn Kanngan ◽  
Vorakan Burapatana ◽  
Warawut Chulalaksananukul
Keyword(s):  
Pichia Pastoris ◽  
Aspergillus Oryzae ◽  
Cloning And Expression ◽  
Beta Glucosidase

scholarly journals Cloning and Expression of an Active Aspartic Proteinase Gene From Aspergillus Oryzae DRDFS13 in Pichia Pastoris

2020 ◽  
Author(s):  
Jermen Mamo ◽  
Konstantina Kostadinovska ◽  
Martin Kangwa ◽  
Marcelo Fernandez-Lahore ◽  
Fassil Assefa
Keyword(s):  
Pichia Pastoris ◽  
Aspergillus Oryzae ◽  
Recombinant Proteins ◽  
Aspartic Protease ◽  
Cloning And Expression ◽  
Major Protein ◽  
Protease Gene ◽  
Recombinant Yeast ◽  
Milk Clotting ◽  
Cheese Production

Abstract BackgroundPichia pastoris is a yeast widely used in expressing recombinant proteins from eukaryotic organisms. In the present study, the total RNA was extracted from a eukaryotic fungus; Aspergillus oryzae DRDFS13 and reverse transcribed into cDNA using specific primers. The gene for aspartic protease was amplified and sequenced and then cloned into pGAPZαA for further expression in P. pastoris. The recombinant yeast (P. patoris X-33Ap) was cultivated in YPD media at pH 5 and 7 for 6 days and the production of recombinant proteins was checked by total protein determination, milk-clotting activity assay, and SDS-PAGE analysis. ResultsThe gene sequence results showed 98% similarity with aspartic protease gene from A. oryzae RIB40. The aspartic protease gene cloned into pGAPZαA (later pMKAP) was successfully expressed in P. pastoris as an active extracellular protease with the highest MCA (190.47 MCU/mL) of secreted enzyme from the recombinant yeast was obtained at pH 5 and 6 days of incubation time. The major protein expressed by the recombinant P. Pastoris X-33 AP has a molecular mass between 32 and 46 kDa. When analyzed for clotting activity, the protein was able to clot skim-milk in 2 min. The clotting activity was found to be 190.47 U/mL.ConclusionThus, the milk-clotting protease extracted form the recombinant yeast in the present study could be a suitable candidate for cheese production. However, further study of the recombinant proteins need to be carried out and its application in cheese production by analyzing the organoleptic and chemical properties of the cheese produced.

Modified pPIC9K vector-mediated expression of a family 11 xylanase gene, Aoxyn11A, from Aspergillus oryzae in Pichia pastoris

2012 ◽  
Vol 63 (3) ◽  
pp. 1109-1120 ◽  
Author(s):  
Jian-Fang Li ◽  
Shu-Juan Gao ◽  
Xiao-Tong Liu ◽  
Yan-Yan Gong ◽  
Zhong-Fa Chen ◽  
...  
Keyword(s):  
Pichia Pastoris ◽  
Aspergillus Oryzae ◽  
Xylanase Gene ◽  
Ppic9k Vector

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).

Aspergillus oryzae S2 AmyA amylase expression in Pichia pastoris: production, purification and novel properties

2018 ◽  
Vol 46 (1) ◽  
pp. 921-932 ◽  
Author(s):  
Sahar Trabelsi ◽  
Mouna Sahnoun ◽  
Fatma Elgharbi ◽  
Rihab Ameri ◽  
Sameh Ben Mabrouk ◽  
...  
Keyword(s):  
Pichia Pastoris ◽  
Aspergillus Oryzae
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