caa a22 4500 60550556X CHVBK 20210128100622.0 cr unu---uuuuu 210128e20151101xx s 000 0 eng 10.1007/s00253-015-6613-2 doi (NATIONALLICENCE)springer-10.1007/s00253-015-6613-2 Cloning, molecular modeling, and docking analysis of alkali-thermostable β-mannanase from Bacillus nealsonii PN-11 [Elektronische Daten] [Prakram Chauhan, Satya Tripathi, Abhays Sangamwar, Neena Puri, Prince Sharma, Naveen Gupta] An alkali-thermostable β-mannanase gene from Bacillus nealsonii PN-11 was cloned by functional screening of E. coli cells transformed with pSMART/HaeIII genomic library. The ORF encoding mannanase consisted of 1100bp, corresponding to protein of 369 amino acids and has a catalytic domain belonging to glycoside hydrolase family 5. Cloned mannanase was smaller in size than the native mannanase by 10kDa. This change in molecular mass could be because of difference in the glycosylation. The tertiary structure of the β-mannanase (MANPN11) was designed and it showed a classical (α/β) TIM-like barrel motif. Active site of MANPN11 was represented by 8 amino acid residues viz., Glu152, Trp189, His217, Tyr219, Glu247, Trp276, Trp285, and Tyr287. Model surface charge of MANPN11 predicted that surface near active site was mostly negative, and the opposite side was positive which might be responsible for the stability of the enzymes at high pH. Stability of MANPN11 at alkaline pH was further supported by the formation of a hydrophobic pocket near active site of the enzyme. To understand the ability of MANPN11 to bind with different substrates, docking studies were performed and found that mannopentose fitted properly into active site and form stable enzyme substrate complex. Springer-Verlag Berlin Heidelberg, 2015 Bacillus nealsonii nationallicence Alkali-thermostable enzyme nationallicence Cloning nationallicence β-Mannanase gene nationallicence Homology modeling nationallicence Docking nationallicence Chauhan Prakram Department of Microbiology, BMS Block, Panjab University South Campus, Chandigarh, India aut Tripathi Satya Department of Pharmacoinformatics, National Institute of Pharmaceutical Science Education and Research, S.A.S, Nagar, Mohali, Punjab, India aut Sangamwar Abhays Department of Pharmacoinformatics, National Institute of Pharmaceutical Science Education and Research, S.A.S, Nagar, Mohali, Punjab, India aut Puri Neena Department of Industrial Microbiology, Guru Nanak Khalsa College, Yamunanagar, Haryana, India aut Sharma Prince Department of Microbiology, BMS Block, Panjab University South Campus, Chandigarh, India aut Gupta Naveen Department of Microbiology, BMS Block, Panjab University South Campus, Chandigarh, India aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/21(2015-11-01), 8917-8925 0175-7598 99:21<8917 2015 99 253 https://doi.org/10.1007/s00253-015-6613-2 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-015-6613-2 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Chauhan Prakram Department of Microbiology, BMS Block, Panjab University South Campus, Chandigarh, India aut NATIONALLICENCE 700 1- Tripathi Satya Department of Pharmacoinformatics, National Institute of Pharmaceutical Science Education and Research, S.A.S, Nagar, Mohali, Punjab, India aut NATIONALLICENCE 700 1- Sangamwar Abhays Department of Pharmacoinformatics, National Institute of Pharmaceutical Science Education and Research, S.A.S, Nagar, Mohali, Punjab, India aut NATIONALLICENCE 700 1- Puri Neena Department of Industrial Microbiology, Guru Nanak Khalsa College, Yamunanagar, Haryana, India aut NATIONALLICENCE 700 1- Sharma Prince Department of Microbiology, BMS Block, Panjab University South Campus, Chandigarh, India aut NATIONALLICENCE 700 1- Gupta Naveen Department of Microbiology, BMS Block, Panjab University South Campus, Chandigarh, India aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/21(2015-11-01), 8917-8925 0175-7598 99:21<8917 2015 99 253