Cloning, expression, purification and characterization of lipase from Bacillus licheniformis, isolated from hot spring of Himachal Pradesh, India

详细信息    查看全文

• 作者：Gagandeep Kaur ; Amninder Singh ; Rohit Sharma ; Vinay Sharma ; Swati Verma…
• 关键词：Lipase ; Bacillus licheniformis ; Metal ions ; Detergents ; Inhibitors ; Solvents
• 刊名：3 Biotech
• 出版年：2016
• 出版时间：December 2016
• 年：2016
• 卷：6
• 期：1
• 全文大小：1,264 KB
• 参考文献：Abramic M, Lescic I, Korica T, Vitale L, Saenger W, Pigac J (1999) Purification and properties of extracellular lipase from Streptomyces rimosus. Enz Microb Technol 25:522–529CrossRef
  Annamalai N, Elayaraja S, Vijayalakshmi S, Balasubramanian T (2011) Thermostable alkaline tolerant lipase from Bacillus licheniformis using peanut oil cake as a substrate. Afr J Biochem Res 5:176–181
  Balan A, Ibrahim D, Rahim RA (2013) Organic-solvent and surfactant tolerant thermostable lipase, isolated from a thermophilic bacterium, Geobacillus thermodenitrificans IBRL-nra. Adv Stud Biol 5:389–401
  Carlsson F, Stalhammar-Carlemalm M, Flardh K, Sandin C, Carlemalm E, Lindahl G (2006) Signal sequence directs localized secretion of bacterial surface proteins. Nature 442:943–946CrossRef
  Castro-Ochoa LD, Rodriguez-Gomez C, Valerio-Alfaro G, Ros RO (2005) Screening, purification and characterization of the thermoalkalophilic lipase produced by Bacillus thermoleovorans CCR11. Enzyme Microb Tech 37:648–654CrossRef
  Chakraborty K, Raj P (2008) An extra-cellular alkaline metallolipase from Bacillus licheniformis MTCC 6824: purification and biochemical characterization. Food Chem 109:727–736CrossRef
  Enujiugha VN, Thani FA, Sanni TM, Abigor RD (2004) Lipase activity in dormant seeds of the african oil bean (Pentaclethra macrophylla Benth). Food Chem 88:405–410CrossRef
  Fullbrook PD (1996) Practical applied kinetics. In: Godfrey T, West S (eds) Industrial enzymology, 2nd edn. Stockholm Press, New York, pp 483–540
  Ghori MI, Iqbal MJ, Hameed A (2011) Characterization of a novel lipase from Bacillus sp. isolated from tannery wastes. Braz J Microbiol 42:22–29CrossRef
  Gohel HR, Ghosh SK, Bragazna VJ (2013) Production, purification and immobilization of extracellular lipases from thermophilic Bacillus subtilis XRF11 and Bacillus licheniformis XRF12 for production of alkyl esters. Int J Lifesci Biotechnol Pharma Res 2:2239–2250
  Goldsack DE (1970) Relation of hydrophobicity index to the thermal stability of homologous proteins. Biopolymers 9:247–252CrossRef
  Gray CJ (1993) Stabilization of enzymes with soluble additives. Thermostability Enzymes 8:124–143
  Guncheva M, Zhiryakova D (2011) Catalytic properties and potential applications of Bacillus lipases. J Mol Catal B Enzym 68:1–25CrossRef
  Gupta R, Gupta N, Rathi P (2004) Bacterial lipases: an overview of production, purification and biochemical properties. Appl Microbiol Biotechnol 64:763–781CrossRef
  Haki GD, Rakshit SK (2003) Developments in industrially important thermostable enzymes: a review. Bioresour Technol 89:17–34CrossRef
  Hayes DG (2004) Enzyme-catalyzed modification of oilseed materials to produce eco-friendly products. J Am Oil Chem Soc 81:1077–1103CrossRef
  Jaeger KE, Eggert T (2002) Lipases for biotechnology. Curr Opin Biotechnol 13:390–397CrossRef
  Kambourova M, Kirilova N, Mandeva R, Derekova A (2003) Purification and properties of thermostable lipase from a thermophilic Bacillus stearothermophilus MC 7. J Mol Catal B Enzym 22:307–313CrossRef
  Kanjanavas P, Khuchareontaworn S, Khawsak P, Pakpitcharoen A, Pothivejkul K, Santiwatanakul S, Matsui K, Kajiwara T, Chansiri K (2010) Purification and characterization of organic solvent and detergent tolerant lipase from Thermotolerant bacillus sp. RN2. Int J Mol Sci 11:3783–3792CrossRef
  Kim MH, Kim HK, Lee JK, Park SY, Oh TK (2000) Thermostable lipase of Bacillus stearothermophilus: high level production, purification, and calcium-dependent thermostability. Biosci Biotechnol Biochem 64:280–286CrossRef
  Kim IH, Kim H, Lee KT, Chung SH, Ko SN (2002) Lipase-catalyzed acidolysis of perilla oil with caprylic acid to produce structured lipids. J Am Oil Chem Soc 79:363–367CrossRef
  Klibanov AM (2001) Improving enzyme by using them in organic solvents. Nature 409:241–246CrossRef
  Kumar S, Kikon K, Upadhyay A, Kanwar SS, Gupta R (2005) Production, purification, and characterization of lipase from thermophilic and alkaliphilic Bacillus coagulans BTS-3. Protein Expr Purif 41:38–44CrossRef
  Lee DW, Koh YS, Kim KJ, Kim BC, Choi HJ, Kim DS, Suhartono MT, Pyun YR (1999) Isolation and characterization of a thermophilic lipase from Bacillus thermoleovorans ID-1. FEMS Microbiol Lett 179:393–400CrossRef
  Lesuisse E, Schanck K, Colson C (1993) Purification and preliminary characterization of the extracellular lipase of Bacillus subtilis 168, an extremely basic pH-tolerant enzyme. Eur J Biochem 216:155–160CrossRef
  Ma J, Zhang Z, Wang B, Kong X, Wang Y, Cao S, Feng Y (2006) Overexpression and characterization of a lipase from Bacillus subtilis. Protein Expr Purif 45:22–29CrossRef
  Mishra B, Madan P (2009) Overexpression, purification and characterization of organic solvent stable lipase from Bacillus licheniformis RSP-09. J Mol Microbiol Biotechnol 17:118–123CrossRef
  Nagao T, Shimada Y, Sugihara A, Murata A, Komemushi S, Tominaga Y (2001) Use of thermostable Fusarium heterosporum lipase for production of structured lipid containing oleic and palmitic acids in organic solvent- free system. J Am Oil Chem Soc 78:167–172CrossRef
  Nawani N, Kaur J (2000) Purification, characterization and thermostability of a lipase from a thermophilic Bacillus sp. J33. Mol Cell Biochem 206:91–96CrossRef
  Nthangeni MB, Patterton HG, van Tonder A, Vergeer WP, Litthauer D (2001) Over-expression and properties of a purified recombinant Bacillus licheniformis lipase: a comparative report on Bacillus lipases. Enzyme Microb Tech 28:705–712CrossRef
  Petersen TN, Brunak S, von Heijne G, Nielson H (2011) Signal P 4.0: discriminating signal peptides from transmembrane regions. Nat Methods 8:785–786CrossRef
  Polizelli PP, Tiera MJ, Bonilla-Rodriguez GO (2008) Effect of surfactants and polyethylene glycol on the activity and stability of a lipase from oilseeds of Pachira aquatica. J Am Oil Chem Soc 85:749–753CrossRef
  Rahman RNZRA, Baharum SN, Basri M, Salleh AB (2005) High yield purification of an organic solvent-tolerant lipase from Pseudomonas sp. Strain S5. Anal Biochem 341:267–274CrossRef
  Rahman RNZRA, Baharum SN, Salleh AB, Basri M (2006) S5 lipase: an organic solvent tolerant enzyme. J Microbiol 44:583–590
  Sanders TH, Pattee HE (1975) Peanut alkaline lipase. Lipids 10:50–54CrossRef
  Sangeetha R, Geetha A, Arulpandi I (2010) Concomitant production of protease and lipase by Bacillus licheniformis VSG1: production, purification and characterization. Braz J Microbiol 41:179–185CrossRef
  Saxena RK, Sheoran A, Giri B, Davidson WS (2003) Purification strategies for microbial lipases. J Microbiol Methods 52:1–18CrossRef
  Schmidt-Dannert C (1999) Recombinant microbial lipases for biotechnological applications. Biorg Med Chem 7:2123–2130CrossRef
  Shariff FM, Rahman RNZRA, Basri M, Salleh AB (2011) A newly isolated thermostable lipase from Bacillus sp. Int J Mol Sci 12:2917–2934CrossRef
  Sharma CK, Kanwar SS (2012) Purification of a novel thermophilic lipase from B. licheniformis MTCC-10498. Int Res J Biol Sci 1:43–48
  Sharma S, Kanwar SS (2014) Organic Solvent Tolerant Lipases and Applications. Sci World J 2014:625258. doi:10.​1155/​2014/​625258
  Sharma R, Soni SK, Vohra RM, Gupta LK, Gupta JK (2002) Purification and characterisation of a thermostable alkaline lipase from a new thermophilic Bacillus sp. RSJ-1. Process Biochem 37:1075–1084CrossRef
  Sharma PK, Singh K, Singh R, Capalash N, Ali A, Mohammad O, Kaur J (2012) Characterization of a thermostable lipase showing loss of secondary structure at ambient temperature. Mol Biol Rep 39:2795–2804CrossRef
  Sigurgisladottir S, Konraosdottir M, Jonsson A, Kristjansson JK, Matthiasson E (1993) Lipase activity of thermophilic bacteria from Icelandic hotsprings. Biotechnol Lett 15:361–366CrossRef
  Wills ED (1960) The relation of metals and –SH groups to the activity of pancreatic lipase. Biochim Biophys Acta 40:481–490CrossRef
  
• 作者单位：Gagandeep Kaur (1)
  Amninder Singh (1)
  Rohit Sharma (2)
  Vinay Sharma (1)
  Swati Verma (3)
  Pushpender K. Sharma (1)
  
  1. Department of Biotechnology, Sri Guru Granth Sahib World University, Fatehgarh Sahib, Punjab, India
  2. NABI, Mohali, Punjab, India
  3. Department of Microbiology, Maharaja Ganga Singh University, Bikaner, Rajasthan, India
  
• 刊物主题：Biotechnology; Agriculture; Cancer Research; Bioinformatics; Stem Cells; Biomaterials;
• 出版者：Springer Berlin Heidelberg
• ISSN：2190-5738

文摘

In the present investigation, a gene encoding extracellular lipase was cloned from a Bacillus licheniformis. The recombinant protein containing His-tag was expressed as inclusion bodies in Esherichia coli BL21DE3 cells, using pET-23a as expression vector. Expressed protein purified from the inclusion bodies demonstrated ~22 kDa protein band on 12 % SDS-PAGE. It exhibited specific activity of 0.49 U mg−1 and % yield of 8.58. Interestingly, the lipase displayed activity at wide range of pH and temperature, i.e., 9.0–14.0 pH and 30–80 °C, respectively. It further demonstrated ~100 % enzyme activity in presence of various organic solvents. Enzyme activity was strongly inhibited in the presence of β-ME. Additionally, the serine and histidine modifiers also inhibited the enzyme activities strongly at all concentrations that suggest their role in the catalytic center. Enzyme could retain its activity in presence of various detergents (Triton X-100, Tween 20, Tween 40, SDS). Sequence and structural analysis employing in silico tools revealed that the lipase contained two highly conserved sequences consisting of ITITGCGNDL and NLYNP, arranged as parallel β-sheet in the core of the 3D structure. The function of these conserve sequences have not fully understood. Keywords Lipase Bacillus licheniformis Metal ions Detergents Inhibitors Solvents