scholarly journals Molecular and biochemical characterization of a novel cold-active and metal ion-tolerant GH10 xylanase from frozen soil

2017 ◽  
Vol 31 (5) ◽  
pp. 955-963 ◽  
Author(s):  
Haiyan Qiu ◽  
Zhongyuan Li ◽  
Hui Wang ◽  
Haiying Zhang ◽  
Shuang Li ◽  
...  
Keyword(s):  
Metal Ion ◽  
Biochemical Characterization ◽  
Frozen Soil ◽  
Cold Active ◽  
Gh10 Xylanase

scholarly journals Purification and biochemical characterization of non-myristoylated recombinant pp60c-src kinase

1992 ◽  
Vol 287 (3) ◽  
pp. 985-993 ◽  
Author(s):  
N B Lydon ◽  
B Gay ◽  
H Mett ◽  
B Murray ◽  
J Liebetanz ◽  
...  
Keyword(s):  
Metal Ion ◽  
Biochemical Characterization ◽  
Src Kinase ◽  
Sf9 Cells ◽  
Peptide Substrates ◽  
Biophysical Study ◽  
Stable Product ◽  
Full Activation

To obtain sufficient material for the biochemical and biophysical study of pp60c-src, we have utilized a recombinant pp60c-src baculovirus lacking the myristoylation site at codon 2. On infection of Sf9 cells, this virus produced large amounts of soluble non-myristoylated pp60c-src. The use of non-myristoylated pp60c-src (1) increases production of pp60c-src compared with the wild-type protein, (2) facilitates purification, (3) yields a stable product and (4) allows biochemical studies in the absence of detergents. Up to 20 mg of pp60c-src of greater than 95% purity has been purified from 6 litres of Sf9 cells grown in a bioreactor. One major and multiple minor forms of pp60c-src were separated by Mono Q f.p.l.c. Isoelectric focusing of purified pp60c-src species revealed heterogeneity, some of which could be attributed to differences in the tyrosine phosphorylation state of the enzyme. Kinetic analysis of non-myristoylated pp60c-src kinase in the presence of Mg2+ gave Km values for angiotensin II and ATP of 2 mM and 30 microM respectively and a Vmax. of 620 nmol/min per mg. The kinetic constants and metal ion preferences of a number of copolymers and peptide substrates have been compared. Polylysine and poly(GLAT), which was not phosphorylated by the pp60c-src kinase, dramatically activated autophosphorylation of Tyr-416, suggesting a conformation modulation of pp60c-src by charged polymers. This finding implies that Tyr-527 dephosphorylation is not sufficient for full activation of pp60c-src in vitro.

scholarly journals BIOCHEMICAL CHARACTERIZATION OF RECOMBINANT PHYTASE ENZYME (phyK) FROM Klebsiella sp. ASR1 ENCAPSULATED WITH ALGINATE

2020 ◽  
Vol 7 (1) ◽  
Author(s):  
Muhammad Eka Hidayatullah ◽  
. Sajidan ◽  
Ari Susilowati ◽  
Baraka Stewart Mkumbe ◽  
Ralf Greiner
Keyword(s):  
Metal Ion ◽  
Biochemical Characterization ◽  
Extreme Conditions ◽  
Optimum Temperature ◽  
High Ph ◽  
Recombinant Phytase ◽  
Ion Effects ◽  
Metal Ion Effects

Karakterisasi Biokimia Enzim Fitase Rekombinan (phyK) dari Klebsiella sp. ASR1 Yang Dienkapsulasi Dengan AlginatEnzim fitase melepas molekul fosfor pada atom C dari benzena Inositol fitat. Tetapi fitase memiliki kelemahan tidak mampu bertahan terhadap kondisi ekstrim dalam lambung nonruminansia. Solusi dalam penelitian ini yaitu fitase dienkapsulasi menggunakan alginat. Penelitian ini bertujuan mengkarakterisasi fitase setelah dienkapsulasi menggunakan alginate. Hasil penelitian ini yaitu fitase yang dienkapsulasi memiliki aktivitas tertinggi pada pH 6,0, sedangkan fitase tanpa enkapsulasi pada pH 5,0. Suhu optimum untuk aktivitas tertinggi fitase yang dienkapsulasi yaitu 70ºC, sedangkan fitase tanpa enkapsulasi 37ºC. Untuk perlakuan penambahan ion logam, aktivitas tertinggi fitase yang dienkapsulasi terjadi dengan penambahan 0,1 mM Fe2+ dan 1,0 mM Ca2+, sedangkan fitase tanpa enkapsulasi dengan penambahan 0,1 mM Fe2+. Berdasarkan hasil penelitian ini, fitase yang dienkapsulasi memiliki keunggulan lebih banyak dibandingkan dengan fitase tanpa enkapsulasi, karena mampu bertahan pada pH dan suhu tinggi, dan beberapa efek ion logam.Kata Kunci: alginat, asam fitat, enkapsulasi, fitase, fitase rekombinanABSTRACTPhytase enzymes release phosphorus molecules on the C atom from benzene inositol phytate. But phytase has the disadvantage of being unable to withstand extreme conditions in the non-ruminant stomach. The solution in this research was phytase encapsulated using alginate. This study aims to characterize phytase after being encapsulated using alginate. The results of this study were the encapsulated phytase had the highest activity at pH 6.0, while the unencapsulated phytase at pH 5.0. The optimum temperature for the highest activity of the encapsulated phytase was 70ºC, while the unencapsulated phytase 37ºC. For treatment of metal ion addition, the highest activity of the encapsulated phytase occurred with the addition of 0.1 mM Fe2+ and 1.0 mM Ca2+, while the unencapsulated phytase with the addition of 0.1 mM Fe2+. Based on the results of this study, the encapsulated phytase had more advantages compared to the unencapsulated phytase, as the former withstand high pH and temperature, and some metal ion effects.

High-level expression and biochemical characterization of a novel cold-active lipase from Rhizomucor endophyticus

2016 ◽  
Vol 38 (12) ◽  
pp. 2127-2135 ◽  
Author(s):  
Xiaojie Duan ◽  
Mingming Zheng ◽  
Yu Liu ◽  
Zhengqiang Jiang ◽  
Shaoqing Yang
Keyword(s):  
Biochemical Characterization ◽  
High Level Expression ◽  
Cold Active ◽  
High Level ◽  
Active Lipase

scholarly journals Biochemical Characterization of Bovine Brain Myristoyl-CoA:Protein N-Myristoyltransferase Type 2

2009 ◽  
Vol 2009 ◽  
pp. 1-9 ◽  
Author(s):  
Ponniah Selvakumar ◽  
Ashakumary Lakshmikuttyamma ◽  
Rajendra K. Sharma
Keyword(s):  
Metal Ion ◽  
Biochemical Characterization ◽  
Kinetic Studies ◽  
Bovine Brain ◽  
Covalent Attachment ◽  
Reading Frame ◽  
Gene Coding ◽  
Acid Protein

Protein N-myristoylation is a lipidic modification which refers to the covalent attachment of myristate, a 14-carbon saturated fatty acid, to the N-terminal glycine residue of a number of mammalian, viral, and fungal proteins. In this paper, we have cloned the gene coding for myristoyl-CoA:protein N-myristoyltransferase (NMT) fromBos tarusbrain. The open reading frame codes for a 410-amino-acid protein and overexpressed inEscherichia coli. Kinetic studies suggested that bovine brain NMT2 and human NMT1 show significant differences in their peptide substrate specificities. The metal ionCa2+had stimulatory effects on NMT2 activity whileMn2+andZn2+inhibited the enzyme activity. In addition, NMT2 activity was inhibited by various organic solvents and other detergents while NMT1 had a stimulatory effect. Biochemical characterization suggested that both forms of NMT have unique characteristics. Further analysis towards functional role NMT2 will lead the development of therapeutic target for the progression of various diseases such as cancer, cardiovascular diseases, and neurodegenerative diseases.

Sign in / Sign up

Export Citation Format

Share Document