Improvement of low-temperature caseinolytic activity of a thermophilic subtilase by directed evolution and site-directed mutagenesis

Biotechnology and Bioengineering

Volumn 104, Issue 5, 2009, Pages 862-870

  Zhong, Chuan Qi ^a   Song, Shengli ^a   Fang, Nan ^a   Liang, Xiaoliang ^a   Zhu, Hui ^a   Tang, Xiao Feng ^a   Tang, Bing ^a  

^a WUHAN UNIVERSITY   (China)

Author keywords

Caseinolytic activity; Cold adaptation; Directed evolution; Thermophilic subtilase

Indexed keywords

AFTER-HEAT; AMINO ACID SUBSTITUTION; CASEINOLYTIC ACTIVITY; CATALYTIC ACTIVITY; COLD ADAPTATION; DIRECTED EVOLUTION; HIGH STABILITY; HIGH-LOW; HOMOLOGY MODELING; HYDROLYTIC ACTIVITIES; INDUSTRIAL USE; LOW TEMPERATURES; LOW-TEMPERATURE ACTIVITY; MULTIPLE ROUTES; OPTIMUM TEMPERATURE; SITE DIRECTED MUTAGENESIS; SUBSTRATE SPECIFICITY; SUBSTRATE-BINDING; SYNTHETIC SUBSTRATES; TEMPERATURE RANGE; THERMOPHILIC SUBTILASE;

AMINES; AMINO ACIDS; ENZYMES; INDUSTRIAL APPLICATIONS; MUTAGENESIS; ORGANIC ACIDS; STRUCTURAL ANALYSIS; SUBSTRATES;

ENZYME ACTIVITY;

PROTEINASE; UNCLASSIFIED DRUG; WF146 PROTEINASE;

AMINO ACID SUBSTITUTION; ARTICLE; BINDING AFFINITY; COLD ACCLIMATIZATION; CONTROLLED STUDY; ENZYME ACTIVITY; GENE MUTATION; LOW TEMPERATURE; NONHUMAN; NUCLEOTIDE SEQUENCE; PROTEIN VARIANT; SITE DIRECTED MUTAGENESIS; TEMPERATURE MEASUREMENT; THERMOSTABILITY;

AMINO ACID SEQUENCE; BACILLUS; BINDING SITES; CASEINS; COLD TEMPERATURE; DIRECTED MOLECULAR EVOLUTION; ENZYME STABILITY; HOT TEMPERATURE; KINETICS; MOLECULAR SEQUENCE DATA; MUTAGENESIS, SITE-DIRECTED; MUTATION, MISSENSE; PROTEIN STRUCTURE, SECONDARY; PROTEIN STRUCTURE, TERTIARY; SUBSTRATE SPECIFICITY; SUBTILISIN;

EID: 70350534535     PISSN: 00063592     EISSN: 10970290     Source Type: Journal    
DOI: 10.1002/bit.22473     Document Type: Article

References (38)

1. Almog O, Gonzalez A, Klein D, Greenblatt HM, Braun S, Shoham G. 2003. The 0.93A crystal structure of sphericase: A calcium-loaded serine protease from Bacillus sphaericus. J Mol Biol 332(5):1071-1082. (Pubitemid 37108800)
2. Almog O, Gonzalez A, Godin N, de Leeuw M, Mekel MJ, Klein D, Braun S, Shoham G, Walter RL. 2009. The crystal structures of the psychrophilic subtilisin S41 and the mesophilic subtilisin Sph reveal the same calcium-loaded state. Proteins 74(2):489-496.
3. Arnold FH, Wintrode PL, Miyazaki K, Gershenson A. 2001. How enzymes adapt: Lessons from directed evolution. Trends Biochem Sci 26(2):100-106. (Pubitemid 32144744)
4. Bian Y, Liang X, Fang N, Tang XF, Tang B, Shen P, Peng Z. 2006. The roles of surface loop insertions and disulfide bond in the stabilization of thermophilic WF146 protease. FEBS Lett 580(25):6007-6014. (Pubitemid 44584299)
5. Bradford MM. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248-254.
6. Bryan PN, Rollence ML, Pantoliano MW, Wood J, Finzel BC, Gilliland GL, Howard AJ, Poulos TL. 1986. Proteases of enhanced stability: Characterization of a thermostable variant of subtilisin. Proteins 1(4):326-334. (Pubitemid 18101960)
7. Cavicchioli R, Siddiqui KS, Andrews D, Sowers KR. 2002. Low-temperature extremophiles and their applications. Curr Opin Biotechnol 13(3):253-261.
8. D'Amico S, Claverie P, Collins T, Georlette D, Gratia E, Hoyoux A, Meuwis MA, Feller G, Gerday C. 2002. Molecular basis of cold adaptation. Philos Trans R Soc Lond B Biol Sci 357(1423):917-925.
9. Davail S, Feller G, Narinx E, Gerday C. 1994. Cold adaptation of proteins. Purification, characterization, and sequence of the heat-labile subtilisin from the antarctic psychrophile Bacillus TA41. J Biol Chem 269(26):17448-17453. (Pubitemid 24218020)
10. DelMar EG, Largman C, Brodrick JW, Geokas MC. 1979. A sensitive new substrate for chymotrypsin. Anal Biochem 99(2):316-320.
11. Eswar N, Ramakrishnan C. 2000. Deterministic features of side-chain mainchain hydrogen bonds in globular protein structures. Protein Eng 13(4):227-238. (Pubitemid 30258956)
12. Feller G, Gerday C. 2003. Psychrophilic enzymes: Hot topics in cold adaptation. Nat Rev Microbiol 1(3):200-208.
13. Guex N, Diemand A, Peitsch MC. 1999. Protein modelling for all. Trends Biochem Sci 24(9):364-367.
14. Jaenicke R. 1981. Enzymes under extremes of physical conditions. Annu Rev Biophys Bioeng 10:1-67.
15. King J, Laemmli UK. 1971. Polypeptides of the tail fibres of bacteriophage T4. J Mol Biol 62(3):465-477.
16. Li SC, Goto NK, Williams KA, Deber CM. 1996. Alpha-helical, but not betasheet, propensity of proline is determined by peptide environment. Proc Natl Acad Sci USA 93(13):6676-6681.
17. Low PS, Bada JL, Somero GN. 1973. Temperature adaptation of enzymes: Roles of the free energy, the enthalpy, and the entropy of activation. Proc Natl Acad Sci USA 70(2):430-432.
18. Miyazaki K, Wintrode PL, Grayling RA, Rubingh DN, Arnold FH. 2000. Directed evolution study of temperature adaptation in a psychrophilic enzyme. J Mol Biol 297(4):1015-1026. (Pubitemid 30692562)
19. Narinx E, Davail S, Feller G, Gerday C. 1992. Nucleotide and derived amino acid sequence of the subtilisin from the antarctic psychrotroph Bacillus TA39. Biochim Biophys Acta 1131(1):111-113.
20. Narinx E, Baise E, Gerday C. 1997. Subtilisin from psychrophilic antarctic bacteria: Characterization and site-directed mutagenesis of residues possibly involved in the adaptation to cold. Protein Eng 10(11):1271-1279. (Pubitemid 28077679)
21. Sarkar G, Sommer SS. 1990. The "megaprimer" method of site-directed mutagenesis. Biotechniques 8(4):404-407.
22. Schwede T, Kopp J, Guex N, Peitsch MC. 2003. SWISS-MODEL: An automated protein homology-modeling server. Nucleic Acids Res 31(13):3381-3385. (Pubitemid 37442164)
23. Siddiqui KS, Cavicchioli R. 2006. Cold-adapted enzymes. Annu Rev Biochem 75:403-433.
24. Siezen RJ, Leunissen JA. 1997. Subtilases: The superfamily of subtilisin-like serine proteases. Protein Sci 6(3):501-523.
25. Strausberg SL, Ruan B, Fisher KE, Alexander PA, Bryan PN. 2005. Directed coevolution of stability and catalytic activity in calcium-free subtilisin. Biochemistry 44(9):3272-3279. (Pubitemid 40321998)
26. Taguchi S, Ozaki A, Momose H. 1998. Engineering of a cold-adapted protease by sequential random mutagenesis and a screening system. Appl Environ Microbiol 64(2):492-495. (Pubitemid 28079810)
27. Taguchi S, Komada S, Momose H. 2000. The complete amino acid substitutions at position 131 that are positively involved in cold adaptation of subtilisin BPN'. Appl Environ Microbiol 66(4):1410-1415. (Pubitemid 30185610)
28. Tanaka T, Matsuzawa H, Ohta T. 1998. Engineering of S2 site of aqualysin I; alteration of P2 specificity by excluding P2 side chain. Biochemistry 37(50):17402-17407. (Pubitemid 29013741)
29. Tanaka S, Matsumura H, Koga Y, Takano K, Kanaya S. 2007. Four new crystal structures of Tk-subtilisin in unautoprocessed, autoprocessed and mature forms: Insight into structural changes during maturation. J Mol Biol 372(4):1055-1069. (Pubitemid 47368354)
30. Volkov AA, Shao Z, Arnold FH. 2000. Random chimeragenesis by heteroduplex recombination. Methods Enzymol 328:456-463.
31. Wati MR, Thanabalu T, Porter AG. 1997. Gene from tropical Bacillus sphaericus encoding a protease closely related to subtilisins from Antarctic bacilli. Biochim Biophys Acta 1352(1):56-62. (Pubitemid 27195964)
32. Wintrode PL, Miyazaki K, Arnold FH. 2000. Cold adaptation of a mesophilic subtilisin-like protease by laboratory evolution. J Biol Chem 275(41):31635-31640. (Pubitemid 30782037)
33. Wright CS, Alden RA, Kraut J. 1969. Structure of subtilisin BPN' at 2.5 angstrom resolution. Nature 221(5177):235-242.
34. Wu J, Bian Y, Tang B, Chen X, Shen P, Peng Z. 2004. Cloning and analysis of WF146 protease, a novel thermophilic subtilisin-like protease with four inserted surface loops. FEMS Microbiol Lett 230(2):251-258. (Pubitemid 38130208)
35. Yang YR, Zhu H, Fang N, Liang X, Zhong CQ, Tang XF, Shen P, Tang B. 2008. Cold-adapted maturation of thermophilic WF146 protease by mimicking the propeptide binding interactions of psychrophilic subtilisin S41. FEBS Lett 582(17):2620-2626.
36. Zecchinon L, Claverie P, Collins T, D'Amico S, Delille D, Feller G, Georlette D, Gratia E, Hoyoux A, Meuwis MA, Sonan G, Gerday C. 2001. Did psychrophilic enzymes really win the challenge? Extremophiles 5(5):313-321.
37. Zhao H, Arnold FH. 1999. Directed evolution converts subtilisin E into a functional equivalent of thermitase. Protein Eng 12(1):47-53. (Pubitemid 29050541)
38. Zhao H, Giver L, Shao Z, Affholter JA, Arnold FH. 1998. Molecular evolution by staggered extension process (StEP) in vitro recombination. Nat Biotechnol 16(3):258-261. (Pubitemid 28120104)