Biochemical characterization and molecular docking of a novel alkaline-stable keratinase from Amycolatopsis sp. BJA-103

被引:1
作者
Yan, Xia [1 ]
Thou, Hanqi [1 ]
Wang, Ruolin [1 ]
Chen, Huan [1 ]
Wen, Bingjie [1 ]
Dong, Mengmeng [1 ]
Xue, Quanhong [2 ]
Jia, Lianghui [1 ]
Yan, Hua [1 ]
机构
[1] Northwest A&F Univ, Coll Life Sci, Yangling 712100, Peoples R China
[2] Northwest A&F Univ, Coll Nat Resources & Environm, Yangling 712100, Peoples R China
关键词
Protease; Amycolatopsis; AlphaFold; Thermostability; Alkaline stability; Molecular docking; Molecular dynamics simulation; MICROBIAL-DEGRADATION; HYDROGEN; WASTE; PURIFICATION; PROTEINASE; INSIGHT;
D O I
10.1016/j.ijbiomac.2025.139669
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Amycolatopsis sp. BJA-103 was isolated for its exceptional feather-degradation capability, leading to the purification, cloning, and heterologous expression of the keratinase enzyme, KER0199. Sequence analysis places KER0199 within the S8 protease family, revealing <60 % sequence similarity to known proteases. The recombinant KER0199-His(6) demonstrates a broad substrate range, along with remarkable thermostability and alkaline stability, exhibiting optimal activity at pH 11.0 and 60 degrees C, despite the absence of cysteine residues essential for disulfide bonding. Structural modeling reveals a predominantly negatively charged surface and a flat, low-electrostatic-potential substrate-binding pocket. Substrate-binding models, predicted using AlphaFold3 and molecular dynamics simulations, indicate that substrates such as casein, chicken feather beta-keratin P2450, and hemoglobin bind to this pocket, forming anti-parallel beta-sheets with residues G97 to G99 and establishing extensive hydrogen bonds with key residues near the enzyme's active site. These findings suggest that AlphaFold-based substrate binding predictions, combined with an analysis of intermolecular forces, provide a valuable tool for assisting in the elucidation of enzyme specificity and substrate recognition. KER0199, the first characterized S8 family keratinase from the Amycolatopsis genus, shows great potential for industrial applications.
引用
收藏
页数:12
相关论文
共 46 条
  • [1] Abdel-Fattah Azza M., 2018, Journal of Genetic Engineering and Biotechnology, V16, P311, DOI 10.1016/j.jgeb.2018.05.005
  • [2] Accurate structure prediction of biomolecular interactions with AlphaFold 3
    Abramson, Josh
    Adler, Jonas
    Dunger, Jack
    Evans, Richard
    Green, Tim
    Pritzel, Alexander
    Ronneberger, Olaf
    Willmore, Lindsay
    Ballard, Andrew J.
    Bambrick, Joshua
    Bodenstein, Sebastian W.
    Evans, David A.
    Hung, Chia-Chun
    O'Neill, Michael
    Reiman, David
    Tunyasuvunakool, Kathryn
    Wu, Zachary
    Zemgulyte, Akvile
    Arvaniti, Eirini
    Beattie, Charles
    Bertolli, Ottavia
    Bridgland, Alex
    Cherepanov, Alexey
    Congreve, Miles
    Cowen-Rivers, Alexander I.
    Cowie, Andrew
    Figurnov, Michael
    Fuchs, Fabian B.
    Gladman, Hannah
    Jain, Rishub
    Khan, Yousuf A.
    Low, Caroline M. R.
    Perlin, Kuba
    Potapenko, Anna
    Savy, Pascal
    Singh, Sukhdeep
    Stecula, Adrian
    Thillaisundaram, Ashok
    Tong, Catherine
    Yakneen, Sergei
    Zhong, Ellen D.
    Zielinski, Michal
    Zidek, Augustin
    Bapst, Victor
    Kohli, Pushmeet
    Jaderberg, Max
    Hassabis, Demis
    Jumper, John M.
    [J]. NATURE, 2024, 630 (8016) : 493 - 500
  • [3] Multifarious revolutionary aspects of microbial keratinases: an efficient green technology for future generation with prospective applications
    Akram, Fatima
    Aqeel, Amna
    Shoaib, Minahil
    ul Haq, Ikram
    Shah, Fatima Iftikhar
    [J]. ENVIRONMENTAL SCIENCE AND POLLUTION RESEARCH, 2022, 29 (58) : 86913 - 86932
  • [4] [Anonymous], 2013, Afr. J. Biotechnol., V12, P19, DOI [10.5897/AJB12.2428, DOI 10.5897/AJB12.2428]
  • [5] Crystal structure of a subtilisin-like serine proteinase from a psychrotrophic Vibrio species reveals structural aspects of cold adaptation
    Arnórsdóttir, J
    Kristjánsson, MM
    Ficner, R
    [J]. FEBS JOURNAL, 2005, 272 (03) : 832 - 845
  • [6] LOW-BARRIER HYDROGEN-BONDS AND ENZYMATIC CATALYSIS
    CLELAND, WW
    KREEVOY, MM
    [J]. SCIENCE, 1994, 264 (5167) : 1887 - 1890
  • [7] Crossover from hydrogen to chemical bonding
    Dereka, Bogdan
    Yu, Qi
    Lewis, Nicholas H. C.
    Carpenter, William B.
    Bowman, Joel M.
    Tokmakoff, Andrei
    [J]. SCIENCE, 2021, 371 (6525) : 160 - +
  • [8] Two novel S1 peptidases from Amycolatopsis keratinophila subsp. keratinophila D2T degrading keratinous slaughterhouse by-products
    Espersen, Roall
    Falco, Francesco C.
    Hagglund, Per
    Gernaey, Krist V.
    Lantz, Anna E.
    Svensson, Birte
    [J]. APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 2020, 104 (06) : 2513 - 2522
  • [9] Subtilisins of Bacillus spp. hydrolyze keratin and allow growth on feathers
    Evans, KL
    Crowder, J
    Miller, ES
    [J]. CANADIAN JOURNAL OF MICROBIOLOGY, 2000, 46 (11) : 1004 - 1011
  • [10] An integrated strategy for the effective production of bristle protein hydrolysate by the keratinolytic filamentous bacterium Amycolatopsis keratiniphila D2
    Falco, Francesco Cristino
    Espersen, Roall
    Svensson, Birte
    Gernaey, Krist, V
    Lantz, Anna Eliasson
    [J]. WASTE MANAGEMENT, 2019, 89 : 94 - 102