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- 저자 : Vikas Beniwal (검색결과 2 건)
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Vinod Chhokar,Vikas Beniwal,Anil Kumar,J. S. Rana,Seema,Raj Kumar Salar,K. S. Nehra 한국생물공학회 2010 Biotechnology and Bioprocess Engineering Vol.15 No.5
A tannase (E.C. 3.1.1.20) producing fungal strain was isolated from soil and identified as Aspergillus heteromorphus MTCC 8818. Maximum tannase production was achieved on Czapek Dox minimal medium containing 1% tannic acid at a pH of 4.5 and 30°C after 48 h incubation. The crude enzyme was purified by ammonium sulfate precipitation and ion exchange chromatography. Diethylaminoethyl-cellulose column chromatography led to an overall purification of 39.74-fold with a yield of 19.29%. Optimum temperature and pH for tannase activity were 50°C and 5.5 respectively. Metal ions such as Ca2+,Fe2+, Cu1+, and Cu2+ increased tannase activity, whereas Hg2+, Na1+, K1+, Zn2+, Ag1+, Mg2+, and Cd2+ acted as enzyme inhibitors. Various organic solvents such as isopropanol,isoamyl alcohol, benzene, methanol, ethanol, toluene, and glycerol also inhibited enzyme activity. Among the surfactants and chelators studied, Tween 20, Tween 80, Triton X-100, EDTA, and 1, 10-o-phenanthrolein inhibited tannase activity, whereas sodium lauryl sulfate enhanced tannase activity at 1% (w/v).
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Identification of PCR-based DNA Marker Linked to High Phytase Level of Wheat
Amit Vashishth,Sewa Ram,Vikas Beniwal 한국작물학회 2018 Journal of crop science and biotechnology Vol.21 No.1
The phytase is a key enzyme to hydrolyze phytic acid present in wheat grains and improves the bio-availability of micronutrients in monogastric animals. Phytase trait being contributed by specific regions of the genome requires identification of these regions, using suitable molecular markers. Hence, in the present investigation we attempted to develop a PCR-based marker that detects the phytase level in wheat. Six sets of PCR primers were designed on the basis of nucleotides sequence variation found in the sequence of both varieties. Out of six set of primers, one set amplified two different sized bands, i.e. 334 bp and 295 in two wheat cultivars C-306 (low phytase) and DBW 17 (high phytase), respectively. It exhibited a polymorphic banding pattern with length polymorphism and clearly separating low and high phytase genotypes. The primer set was also used for PCR of 46 synthetic hexaploids and 46 release varieties of wheat to validate the developed markers. Association among identified markers and phytase activity was found to be at 99.9% confidence level based on Fisher’s exact test (F-test). Therefore, this PCR primer set will be useful to select the wheat germplasm having high phytase levels and also in wheat breeding programs aimed at improving phytase levels in bread wheat cultivars.
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