Random mutagenesis and process optimization of bacterial co-culture for hyperproduction of 1, 4-α-D-glucan glucanohydrolase using submerged fermentation
Article Sidebar
Main Article Content
Abstract
The exponential increase in the application of 1,4-α-D-glucan glucanohydrolase (GGH) in various fields has placed stress and demand in both qualitative improvement and quantitative enhancement through strain improvement. In the present work, Bacillus subtilis LCBT-15 and Bacillus amyloliquefaciens LCBT-20 were subjected to physical as well as chemical mutagenesis for improving the GGH production potential. Applications of the UV light and ethidium bromide did not cause a significant increase in the enzyme production. However, Ethyl methane sulphonate (EMS) treated co-culture 10 gave 1.3-fold increase in the GGH production, in contrast to the wild co-culture. Different physicochemical parameters including fermentation media, rate of fermentation, temperature, pH, nitrogen and carbon sources and surfactants were also investigated. The M7 medium composition was optimized for GGH production after 48h of incubation at 37oC and pH 6. The optimum inoculum size was 3.5 ml (1´106 cells/ml) in 50 ml of medium. The best carbon and nitrogen sources were lactose (2.5 %); ammonium chloride (1.75 %) and beef extract (1 %), respectively. Optimal GGH production (287 U/ml) was obtained when the medium was supplemented with 0.05% Tween 80. The novelty of this work was exploration of the synergistic phenomena of mutant bacterial co-culture for the enhancement of GGH production.
Downloads
Article Details
Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Creative Commons Attribution License that allows others to share the work with an acknowledgement of the work's authorship and initial publication in this journal.
Authors grant to the Publisher the following rights to the manuscript, including any supplemental material, and any parts, extracts or elements thereof:
- the right to reproduce and distribute the Manuscript in printed form, including print-on-demand;
- the right to produce prepublications, reprints, and special editions of the Manuscript;
- the right to translate the Manuscript into other languages;
- the right to reproduce the Manuscript using photomechanical or similar means including, but not limited to photocopy, and the right to distribute these reproductions;
- the right to reproduce and distribute the Manuscript electronically or optically on any and all data carriers or storage media – especially in machine readable/digitalized form on data carriers such as hard drive, CD-Rom, DVD, Blu-ray Disc (BD), Mini-Disk, data tape – and the right to reproduce and distribute the Article via these data carriers;
- the right to store the Manuscript in databases, including online databases, and the right of transmission of the Manuscript in all technical systems and modes;
- the right to make the Manuscript available to the public or to closed user groups on individual demand, for use on monitors or other readers (including e-books), and in printable form for the user, either via the internet, other online services, or via internal or external networks.
References
Haq I, Javed MM, Hameed U, Adnan F. Kinetics and thermodynamic studies of alpha-amylase from Bacillus licheniformis mutant. Pak J Bot. 2010; 45: 3507-3516.
Paul D. Microorganisms and α-amylase: a concise review. Inov J Life Sci. 2016; 4, 1-5.
Dehkordi MM, Javan FA. Application of alpha-amylase in biotechnology. J Biol Today's World. 2012; 1: 39-50.
Tiwari SP, Srivastava R, Singh CS, Shukla K, Singh RK, Singh P, et al. Amylases: an overview with special reference to alpha amylase. J Global Biosci. 2015; 4: 1886-1901.
Goers L, Freemont P, Polizzi KM. Co-culture systems and technologies: taking synthetic biology to the next level. J R Soc Interface. 2014; 17: 1-13.
Hesseltine CW. Mixed-culture fermentations, 1st; National Academies Press (US): Washington, 1992; pp. 52-57.
Mithun S, Dipak V. Mutagenesis of Lactobacillus species for the generation of a mutant with hyper-producing dextransucrase activity. World J Pharm Res. 2015; 5: 777-786.
Szafraniec K, Wloch DM, Sliwa P, Borts RH, Korona R. Small fitness effects and weak genetic interactions between deleterious mutations in heterozygous loci of the yeast Saccharomyces cerevisiae. Genet Res Camb. 2003; 82: 19-31.
Ho HL, Chor XK. Improvement of xylanase production by Bacillus subtilis in submerged fermentation after UV and chemicals mutagenesis. J Adv Biol Biotechnol. 2015; 3: 42-57.
Francis F, Sabu A, Nampoothiri KM, Ramachandran S, Ghosh S, Szakacs G, Pandey A.Use of response surface methodology for optimizing process parameters for the production of α-amylase by Aspergillus oryzae. Biochem Eng J. 2003; 15: 107-115.
Gangadharan D, Sivaramakrishnan S, Nampoothiri KM, Sukumaran RK, Pandey A. Response surface methodology for the optimization of alpha amylase production by Bacillus amyloliquefaciens. Bioresour Technol. 2008; 99: 4597-4602.
Salman T, Kamal M, Ahmed M, Siddiqa SM, Khan RA, Hassan A. Medium optimization for the production of amylase by Bacillus subtilis RM16 in shake-flask fermentation. Pak J Pharm Sci. 2016; 29: 439-444.
Abdullah R, Naeem N, Aftab M, Kaleem A, Iqtedar M, Iftikhar, T , Naz S. Enhanced production of alpha amylase by exploiting novel bacterial co-culture technique employing solid state fermentation. Iran J Sci Technolo Trans Sci. 2018; 42: 305-318.
Haq I, Ali S, Saleem A, Javed MM. Mutagenesis of Bacillus licheniformis through ethyl methane sulfonate for alpha amylase production. Pak J Bot. 2009; 41: 1489-1498
Riaz A, Qadar S, Anwar A, Iqbal S, Bano S. Production and characterization of thermostable α-amylase from a newly isolated strain of Bacillus subtilis KIBGE-HAR. J Microbiol. 2009; 6: 1-8.
Raj V, Hemashenpagam N. Production and medium optimization of amylase by Bacillus using fermentation methods. J Microbiol Biotechnol. 2012; 2: 481-484.
Akcan N. High Level Production of Extracellular α-amylase from Bacillus licheniformis ATCC 12759 in submerged fermentation. Rom Biotechnol Lett. 2011; 16: 6833-6840.
Vengadaramana A, Balakumar S, Arasaratnam V. Optimization of fermentation medium components to improve α-amylase production by submerged fermentation technology. Sch Acad J Pharm. 2013; 2: 180-186.
Haq I, Ali S, Javed MM, Hameed U, Saleem, A, Adnan F, Qadeer MA. Production of alpha amylase from a randomly induced mutant strain of Bacillus amyloliquefaciens and its application as a desizer in textile industry. Pak J Bot. 2010; 42: 473-484.
Miller GL. Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal Chem. 1959; 31: 426-428.
Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976; 72: 248-254.
Li E, Orduña RMD. A rapid method for the determination of microbial biomass by dry weight using a moisture analyser with an infrared heating source and an analytical balance. Lett Appl Microbiol. 2010; 50: 283-288.
Siddique S, Syed Q, Adnan A, Qureshi FA. Production and screening of high yield avermectin B1b mutant of Streptomyces avermitilis 41445 through mutagenesis. Jandishpur J Microbiol. 2014; 7: 1-8.
Suribabu K, Govardhan TL, Hemalatha KPJ. Strain improvement of Brevibacillus borostelensis R1 for optimization of α-Amylase production by mutagens. J Microb Biochem Technol. 2014; 6: 123-127.
Haq I, Ali S, Saleem A, Javed MM. Mutagenesis of Bacillus licheniformis through ethyl methane sulfonate for alpha amylase production. Pak J Bot. 2009; 41: 1489-1498.
Malhotra R, Noorwez SM, Satyanarayana T. Production and partial characterization of thermostable and calcium-independent α-amylase of an extreme thermophile Bacillus thermooleovorans NP54. Lett Appl Microbiol. 2000; 31: 378-384.
Suribabu K, Govardhan T L, Hemalatha K. Optimization of various nitrogen sources for the production of amylase using Brevibacillus borstelensis R1 by submerged fermentation. Int J Curr Microbiol Appl Sci. 2014; 3 : 791-800.
Mahmood S, Shahid MG, Nadeem M, Irfan M, Syed Q. Production and optimization of alpha amylase from Apergillus niger using potato peel as substrate. Pak J Biotechnol. 2016; 13: 101-109.
Punia P, Kaushik S, Jyoti A. Optimization of production conditions and partial characterization of extracellular amylase from Bacillus Subtilis under submerged condition. J Sci Ind Res. 2016; 75: 371-377.
Riaz N, Haq I, Qadeer MA. Characterization of α-Amylase by Bacillus subtilis. Int J Agric Biol. 2003; 5: 249–252.
Cheba B, Zaghloul T, Isolation and identification of a marine bacterium and optimization of its amylase production. J Appl Biol Biotechnol. 2008; 1: 29-33.
Vijayabaskar P, Jayalakshmi D, Shankar T. Amylase production by moderately halophilic Bacillus cereus in solid state fermentation. Afr J Microbiol Res. 2012; 6: 4918-4926.
Mahalakshmi N, Jayalakshmi S. Amylase, cellulase and xylanase production from a novel bacterial isolate Achromobacter xylosoxidans isolated from marine environment. Int J Adv Res Biol Sci. 2016; 3: 230-233.
Abdullah R, Ali S, Aslam A, Haq I. Influence of different carbon sources on the production of alpha amylase by Aspergillus oryzae on kinetic basis. Pak J Biotechnol. 2005; 2: 89-94.
Dash BK, Rahman MM, Sarker PK. Molecular identification of a newly isolated Bacillus subtilis BI19 and optimization of production conditions for enhanced production of extracellular amylase. BioMed Res Int. 2015; 1: 1-9
Swain MR, Kar S, Padmaja G, Ray RC. Partial characterization and optimization of production of extracellular alpha amylase from Bacillus subtilis isolated from culturable cow dung microflora. J Microbiol. 2006; 55: 289-296.
Saxena L, Iyer BK, Ananthanarayan L. Three phase partitioning as a novel method for purification of ragi (Eleusine coracana) bifunctional amylase/protease inhibitor. Process Biochem 2007; 42: 491-495.
Simair AA, Qureshi AS, Khushk I, Ali CH, Lashari S, Bhutto MA, et al. Production and partial characterization of α-amylase enzyme from Bacillus sp. BCC 01-50 and potential applications. BioMed Res Int. 2017; 1: 1-9.
Sivakumar T, Shankar T, Vijayabaskar P, Muthukumar J, Nagendrakannan E. amylase production using Bacillus cereus isolated from a vermicompost site. Intl J Microbiol Res. 2012; 3: 117-123.