Screening of Lipase-Producing Bacterial Strains and Analysis of Enzyme Production Characteristics

¹ College of Health Management, Shanghai Jian Qiao University, Shanghai 201306, China; School of Life Sciences, Shanghai University, Shanghai 200444, China,

² School of Life Sciences, Shanghai University, Shanghai 200444, China,

³ Junior High School Department, The Affiliated School of Shanghai University, Shanghai 200444, China,

IJGPN 2025 , 3(0), 149–158; https://doi.org/10.26689/ijgpn.v3iSpecial.13936

© 2025 by the Authors. Licensee Whioce Publishing, Singapore. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution -Noncommercial 4.0 International License (CC BY-NC 4.0) ( https://creativecommons.org/licenses/by-nc/4.0/ )

Abstract

Lipase, as an important biological catalyst, hydrolyzes triglycerides into fatty acids, diglycerides, monoglycerides, and glycerol, and is widely used in food, leather, pharmaceutical, and daily chemical industries, particularly in the food sector. Lipase is abundantly present in microorganisms and animals, with lipase-producing microorganisms being suitable for large-scale industrial production due to their rapid reproduction, short cycles, and diverse species. However, since there are currently few strains capable of producing high lipase activity, the increasing demand for industrial applications has made the search for more novel lipase-producing strains increasingly crucial. In this study, from 15 bacterial strains initially isolated from soil in the laboratory, one lipase-producing strain, zjy1-5, was identified through preliminary screening on glycerol-containing plates. Through 16S rDNA sequence analysis and Gram staining microscopic observation, the strain was identified as Rhodococcus rhodochrous, a Gram-positive bacterium. Subsequently, the fermentation conditions for lipase production were optimized, revealing that glucose was the optimal carbon source and 37°C the optimal culture temperature for lipase production. This research provides an experimental basis for further development and utilization of bacterial lipase.

References

Joseph B, Ramteke PW, Thomas G, 2008, Cold Active Microbial Lipases: Some Hot Issues and Recent Developments. Biotechnol Adv, 26(5): 457–470.

Davis BG, Boyer V, 2001, Biocatalysis and Enzymes in Organic Synthesis. Nat Prod Rep, 18: 618–640.

Wang L, Cai Y, Liao X, et al., 2008, Optimization of Fermentation Conditions for a Low-Temperature Lipase-Producing Strain. J Food Sci Biotechnol, 27(3): 114–119.

Niu D, Zhang Y, 2002, Selection of an Alkaline Lipase-Producing Strains and Optimalization of Its Fermentation Conditions. Food Ferment Ind, 29(5): 28–31.

Li J, Shen W, Fan G, et al., 2018, Screening, Purification and Characterization of Lipase from Burkholderia pyrrocinia B1213. 3 Biotech, 8(9): 387.

Ramani K, Chockalingam E, Sekaran G, 2010, Production of a Novel Extracellular Acidic Lipase from Pseudomonas gessardii Using Slaughterhouse Waste as a Substrate. J Ind Microbiol Biotechnol, 37(5): 531–535.

Kantak JB, Bagade AV, Mahajan SA, et al., 2011, Isolation, Identification and Optimization of a New Extracellular Lipase-Producing Strain of Rhizopus sp. Appl Biochem Biotechnol, 164(7): 969–978.

Rasol R, Rashidah AR, Nazuha RS, et al., 2014, Psychrotrophic Lipase Producers from Arctic Soil and Sediment Samples. Pol J Microbiol, 63(1): 75–82.

Kiplimo D, Mugweru J, Kituyi S, et al., 2019, Diversity of Esterase and Lipase-Producing Haloalkaliphilic Bacteria from Lake Magadi in Kenya. J Basic Microbiol, 59(12): 1173–1184.

Dahiya P, Arora P, Chaudhury A, et al., 2010, Characterization of an Extracellular Alkaline Lipase from Pseudomonas mendocina M-37. J Basic Microbiol, 50(5): 420–426.

Li Z, Tang X, Huang W, et al., 2011, Rheology, Microstructure, and Baking Characteristics of Frozen Dough Containing Rhizopus chinensis Lipase and Transglutaminase. Cereal Chem, 88(6): 596–601.

Cho SS, Park DJ, Simkhada JR, et al., 2012, A Neutral Lipase Applicable in Biodiesel Production from a Newly Isolated Streptomyces sp. CS326. Bioprocess Biosyst Eng, 35(1–2): 227–234.

Cherif S, Mnif S, Hadrich F, et al., 2011, A Newly High Alkaline Lipase: An Ideal Choice for Application in Detergent Formulations. Lipids Health Dis, 10: 221.

Rashid N, Shimada Y, Ezaki S, et al., 2001, Low-Temperature Lipase from Psychrotrophic Pseudomonas sp. Strain KB700A. Appl Environ Microbiol, 67(9): 4064–4069.

Zhang B, Yang J, Shu H, et al., 2007, Screening, Identification of Lipase Production Bacteria and Optimization of the Lipase Production. Biotechnology, 17(1): 23–26.

Guo W, Guo X, Yuan H, et al., 2013, Screening and Identification of Efficiently-Producing Fungi from Lipase and Optimization of Its Lipase-Producing Condition. Feed Ind, 34(4): 39–44.

Mo Q, Liu A, Guo H, et al., 2016, A Novel Thermostable and Organic Solvent-Tolerant Lipase from Xanthomonas oryzae pv. oryzae YB103: Screening, Purification and Characterization. Extremophiles, 20(2): 157–165.

Das A, Shivakumar S, Bhattacharya S, et al., 2016, Purification and Characterization of a Surfactant-Compatible Lipase from Aspergillus tamarii JGIF06 Exhibiting Energy-Efficient Removal of Oil Stains from Polycotton Fabric. 3 Biotech, 6(2): 1–8.

Ayaz B, Ugur A, Boran R, 2015, Purification and Characterization of Organic Solvent-Tolerant Lipase from Streptomyces sp. OC119-7 for Biodiesel Production. Biocatal Agric Biotechnol, 4(1): 103–108.

Ungcharoenwiwat P, H-kittikun A, 2015, Purification and Characterization of Lipase from Burkholderia sp. EQ3 Isolated from Wastewater from a Canned Fish Factory and Its Application for the Synthesis of Wax Esters. J Mol Catal B Enzym, 115: 96–104.

Wen J, Shi B, Wu W, et al., 2008, Isolation and Identification of a Lipase-Producing Strain of Bacillus subtilis FS321 and Its Preliminary Optimization for Lipase Production. Food Ferment Ind, 34(2): 37–41.

Shang H, 2009, Screening and Molecular Identification of Lipase-Producing Strain. J Hebei Agri Sci, 13(10): 50–52, 81.

Gao H, Wu G, (eds), 2015, Laboratory Manual for Microbiology, Higher Education Press, Beijing, China, 29–30.

Moreno C, Romero J, Espejo RT, 2002, Polymorphism in Repeated 16S rRNA Genes is a Common Property of Type Strains and Environmental Isolates of the Genus Vibrio. Microbiology (Reading), 148: 1233–1239.

Huo Y, Danecka W, Farquhar I, et al., 2025, The Type of Carbon Source Not the Growth Rate It Supports Can Determine Diauxie in Saccharomyces cerevisiae. Commun Biol, 8: 325.

Kantak JB, Bagade AV, Mahajan SA, et al., 2011, Isolation, Identification and Optimization of a New Extracellular Lipase-Producing Strain of Rhizopus sp. Appl Biochem Biotechnol, 164(7): 969–978.

Dalmau E, Montesinos JL, Lotti M, et al., 2000, Effect of Different Carbon Sources on Lipase Production by Candida rugosa. Enzyme Microb Technol, 26(9–10): 657–663.

Nguyen K, Lee CW, 2025, Catalytic His-Loop Flexibility Drives High Activity in Hyperthermophilic Esterase EstE1 While Preserving Structural Stability. Microbiol Spectr, 13(10): e0100325.

Kumar S, Kikon K, Upadhyay A, et al., 2005, Production, Purification, and Characterization of Lipase from Thermophilic and Alkaliphilic Bacillus coagulans BTS-3. Protein Expr Purif, 41(1): 38–44.

Qiao H, Zhang W, Guan W, et al., 2017, Enhanced Expression of Lipase I from Galactomyces geotrichum by Codon Optimisation in Pichia pastoris. Protein Expr Purif, 138: 34–45.

Liu G, Hu S, Li L, et al., 2015, Purification and Characterization of a Lipase with High Thermostability and Polar Organic Solvent-Tolerance from Aspergillus niger AN0512. Lipids, 50(11): 1155–1163.

Tripathi R, Singh J, Bharti RK, et al., 2014, Isolation, Purification and Characterization of Lipase from Microbacterium sp. and Its Application in Biodiesel Production. Energy Procedia, 54: 518–529.

Bae JH, Kwon MH, Kim IH, et al., 2014, Purification and Characterization of a Cold-Active Lipase from Pichia lynferdii Y-7723: pH-Dependent Activity Deviation. Biotechnol Bioprocess Eng, 19(5): 851–885.

Shu Z, Lin H, Shi S, et al., 2016, Cell-Bound Lipases from Burkholderia sp. ZYB002: Gene Sequence Analysis, Expression, Enzymatic Characterization, and 3D Structural Model. BMC Biotechnol, 16(1): 38.

Miao C, Luo W, Lv P, et al., 2014, Screening, Identification of the Lipase-Producing Strains and Optimization of the Lipase Production. Acta Energiae Solaris Sinica, 35(9): 1708–1714.

Bora L, 2014, Purification and Characterization of Highly Alkaline Lipase from Bacillus licheniformis MTCC 2465: A Study of Its Detergent Compatibility and Applicability. J Surfactants Deterg, 17(5): 889–898.

Hong Y, Shi P, Lu H, et al., 2015, A Thermophilic β-Mannanase from Neosartorya fischeri P1 with Broad pH Stability and Significant Hydrolysis Ability of Various Mannan Polymers. Food Chem, 173(173): 283–289.

Aulitto M, Fusco S, Limauro D, et al., 2019, Galactomannan Degradation by Thermophilic Enzymes: A Hot Topic for Biotechnological Applications. World J Microbiol Biotechnol, 35(2): 32.

Sadaqat B, Dar MA, Sha C, et al., 2024, Thermophilic β-Mannanases from Bacteria: Production, Resources, Structural Features and Bioengineering Strategies. World J Microbiol Biotechnol, 40(4): 130.

Chen X, Tian Z, Zhou H, et al., 2023, Enhanced Enzymatic Performance of β-Mannanase Immobilized on Calcium Alginate Beads for the Generation of Mannan Oligosaccharides. Foods, 12(16): 3089.

Sun D, Li C, Cui P, et al., 2022, Reshaping the Binding Channel of a Novel GH113 Family β-Mannanase from Paenibacillus cineris (PcMan113) for Enhanced Activity. Bioresour Bioprocess, 9(1): 17.

Ahirwar S, Soni H, Rawat HK, et al., 2016, Experimental Design of Response Surface Methodology Used for Utilisation of Palm Kernel Cake as Solid Substrate for Optimised Production of Fungal Mannanase. Mycology, 7(3): 143–153.

Li YX, Wang NN, Yan QJ, et al., 2022, A Novel Neutral Thermophilic β-Mannanase from Malbranchea cinnamomea for Controllable Production of Partially Hydrolyzed Konjac Powder. Appl Microbiol Biotechnol, 106(5–6): 1919–1932.

Previous article in this issue

Next article in this issue