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Cloning of a novel phytase from an anaerobic rumen bacterium, Mitsuokella jalaludinii, and its expression in Escherichia coli

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Journal of Integrative Agriculture 2015年第9期14卷 1816-1826页

作者： TAN Wan-qin Phang Chiun Yee Sieo Chin Chin Yiap Beow Chin clemente michael wong vui ling Norhani Abdullah Son Radu Ho Yin Wan Laboratory of Vaccines and Immunotherapeutics Institute of BioscienceUniversiti Putra Malaysia Department of Microbiology Faculty of Biotechnology and Biomolecular SciencesUniversiti Putra Malaysia Department of Life Sciences School of Pharmacy and HealthFaculty of Medicine and HealthInternational Medical University Biotechnology Research Institute Universiti Malaysia Sabah Institute of Tropical Agriculture Universiti Putra Malaysia Department of Biochemistry Faculty of Biotechnology and Biomolecular SciencesUniversiti Putra Malaysia Department of Food Science Faculty of Food Science and TechnologyUniversiti Putra Malaysia

The ful length phytase gene of Mitsuokel a jalaludini was successful y cloned and was found to be 1 047 bp in length, with 348 amino acids, and was designated as PHY7 phytase gene. A comparison of the sequence of PHY7 phytase gene of M. jalaludini with various microbial phytase gene sequences showed that it was not similar to those from other bacteria except Selenomonas ruminatium, thus suggesting that they may both express a new class of phytase. The PHY7 phytase gene was subsequently subcloned into bacterial expression vector, pET32a, for expression in Escherichia coli strain Ro-setta-gami. Expression of the recombinant phytase gene was optimised and characterised. The recombinant phytase was estimated to be approximately 55 kDa by SDS-PAGE analysis. The recombinant phytase exhibited optimum activity at 55°C, pH 4.5 and showed good pH stability from pH 3.5 to 5.5 (>78%relative activity). Metal ions such as Ca2+, Mg2+, and K+were found to exert signiifcant stimulatory effect on the recombinant phytase activity while Cu2+, Fe3+, and Zn2+greatly inhibited the enzyme activity. The recombinant phytase showed moderate resistance to trypsin proteolysis, but susceptible to pepsin proteolysis. The results of the study showed that several characteristics of recombinant phytase were slightly different from the native enzyme. Unfavourable characteristics such as reduced pH stability and metal ion effects should be taken into consideration during feed enzyme formulation.

关键词： phytase cloning recombinant Escherichia coli Mitsuokella jalaludinii

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A comparative analysis of β-mannanases of bacteria from Antarctica and Malaysia

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Advances in Polar Science 2013年第4期24卷 202-207页

作者： clemente michael vui ling wong Heng Keat Tam Hui Yee Yong Biotechnology Research Institute Universiti Malaysia SabahJalan UMS

β-mannanase is an enzyme that is commonly expressed in environmental bacteria. It degrades hemicellulose found in plant material and recycles nutrients back into the environment. Because this enzyme significantly contributes to biodegradation and has recently been applied in industry, we conducted a comparative analysis of bacterial isolates found in soil samples from Schirmacher Oasis, Antarctica, and Sabah, Malaysia that were capable of degrading mannan. A total of 9 bacterial isolates from Antarctica and 30 bacterial isolates from Malaysia exhibited β-mannanase activity. These bacteria were differentiated and clustered using their random amplified polymorphic DNA （RAPD） profiles, and the β-mannanase activity of these isolates was tested at different temperatures and pH. Five out of 9 Antarctica isolates and seven out of 30 Malaysian isolates were identified based on their 16S rDNA sequences. Identified bacterial isolates from Antarctica were： MP1 （Bacillus amyloliquefaciens）, MP2 （Bacillus pumilus）, MP5 （Bacilluspumilus）, A40 （Arthrobacter sp.）, and C27 （Arthrobacter oxydans）. Identified bacterial isolates from Ma- laysia were： Y1 （Paenibacillus sp.）, Y2 （Bacillus sp.）, Y16 （Paenibacillus sp.）, Y18 （Paenibacillus sp.）, A7 （Paenibacillus sp.）, B26 （Streptomyces sp.）, and D4 （Paenibacillus amylolyticus）. β-mannanases produced by the Antarctica bacterial isolates MP1 （Bacillus amyloliquefaciens） and A40 （Arthrobacter sp.） were active at 5℃ and 20℃, respectively, while the β-mannanase pro- duced by the bacterial isolate from Malaysia, A7 （Paenibacillus sp.）, was active at 35 ℃.

关键词： Antarctic regions guar gum locust bean gum hemicellulose and hemicellulase

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Genome of a thermophilic bacterium Geobacillus sp. TFV3 from Deception Island, Antarctica

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Advances in Polar Science 2020年第2期31卷 146-152页

作者： Xin Jie CHING Chui Peng TEOH Dexter J.H.LEE Marcelo GONZáLEZ-ARAVENA Nazalan NAJIMUDIN Yoke Kqueen CHEAH Paris LAVIN clemente michael vui ling wong Biotechnology Research Institute Universiti Malaysia SabahJalan UMS88400 Kota KinabaluSabahMalaysia Instituto Antártico Chileno Plaza Munoz Gamero 1055Punta ArenasChile School of Biological Science Universiti Sains MalaysiaPersiaran Bukit Jambul 11900 Bayan LepasPenangMalaysia Department of Biomedical Science Faculty of Medicine and Health SciencesUniversiti Putra Malaysia43400 UPM SerdangSelangor Darul EhsanMalaysia Universidad de Antofagasta Facultad de Ciencias del Mary Recursos BiologicosDepartamento de BiotecnologiaLaboratorio de Complejidad Microbianay Ecología FuncionalInstituto AntofagastaChile National Antarctic Research Centre University of Malaya50603 Kuala LumpurMalaysia

Thermophilic microorganisms have always been an important part of the ecosystem,particularly in a hot environment,as they play a key role in nutrient recycling at high temperatures where most microorganisms cannot *** most of the thermophiles are archaea,thermophiles can also be found among some species of *** bacteria are very useful in the fundamental study of heat adaptation,and they are also important as potential sources of thermostable enzymes and ***,we have isolated a Gram-positive thermophilic bacterium,Geobacillus ***3 from a volcanic soil sample from Deception Island,*** project was undertaken to analyze the genes of this thermophilic Antarctic bacterium and to determine the presence of thermal-stress adaptation proteins in its *** genome of Geobacillus ***3 was first purified,sequenced,assembled,and *** complete genome was found to harbor genes encoding for useful thermal-stress adaptation *** majority of these proteins were categorized under the family of molecular chaperone and heat shock *** genomic information could eventually provide insights on how the bacterium adapts itself towards high growth temperatures.

关键词： temperature 16SrDNA genomes Geobacillus Deception Island

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