Advertisement

3 Biotech

, 9:396 | Cite as

Production and purification of laccase by Bacillus sp. using millet husks and its pesticide degradation application

  • P. Srinivasan
  • T. Selvankumar
  • S. Kamala-Kannan
  • R. Mythili
  • A. Sengottaiyan
  • M. GovarthananEmail author
  • B. Senthilkumar
  • K. SelvamEmail author
Short Reports
  • 11 Downloads

Abstract

Lignocellulosic agricultural bi-products, pearl millet (PM) and finger millet (FM) husks, were used for the production of laccase using Bacillus sp. PS under solid-state fermentation (SSF). Abiotic variables such as substrate (PM, FM) concentration (1–5%), incubation time (24–96 h) and pH (5–10) were optimized using Response surface methodology (RSM) to maximize the laccase production. The predicted model showed maximum laccase activity of 402 U/mL appearing after 96 h of incubation with PM 2.0 g/L and FM 1.5 g/L at pH 7.0. Single protein band on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) confirmed homogeneity of the laccase with a molecular weight of 63–75 kDa. The partially purified laccase effectively degraded the pesticides (Tricel, 71.8 ± 3.5 and Phoskill 77.3 ± 3.4%) within 5 days of incubation (40 °C) in pH 7.0. The pesticide degradation was further confirmed by high-performance liquid chromatography (HPLC) and the chromatograms showed the single dominant peaks at retention time 2.482 (tricel) and 2.608 (phoskill) min, respectively. Pesticide-degrading laccase was produced by Bacillus sp. PS under SSF reveals the utilization of low-cost bi-substrates for enhanced laccase production.

Keywords

Bacillus sp. Finger millet Pearl millet Laccase Phoskill Tricel 

Notes

Acknowledgements

This work is partially supported by the Department of Biotechnology, Mahendra Arts and Science College (Autonomous), and the Department of Science and Technology, Government of India (DST-FIST sponsored—Ref. No. SR/FST/College-232/2014).

Compliance with ethical standards

Conflict of interest

The authors declare that they do not have any conflict of interest.

References

  1. Amitai G, Adani R, Sod-Moriah G, Rabinovitz I, Vincze A, Leader H, Chefetz B, Leibovitz-Persky L, Friesem D, Hadar Y (1998) Oxidative biodegradation of phosphorothiolates by fungal laccase. FEBS Lett 438:195–200CrossRefGoogle Scholar
  2. Audu SS, Ehanwo AA, Aremu MO, Tukura BW, Ambo AI, Usman A (2018) Chemical composition of finger millet (Eleusine coracana). Trends Food Sci Tech 3(2):905–908Google Scholar
  3. Azhari AMN, Adiamo OQ, Awad RM, Babiker EE (2017) Changes in chemical composition and total energy as affected by fermentation and/or cooking of pearl millet flour supplemented with Moringa or fenugreek seeds flour. Food Res Int 24(4):1562–1570Google Scholar
  4. Binod P, Satyanagalakshmi K, Sindhu R, Janu KU, Sukumaran RK, Pandey A (2012) Dilute acid pretreatment and enzymatic saccharification of sugarcane tops for bioethanol production. Renew Energy 37:109–116CrossRefGoogle Scholar
  5. Chauhan PS, Goradia B, Saxena A (2017) Bacterial laccase: recent update on production, properties and industrial applications. 3 Biotech 7:323CrossRefGoogle Scholar
  6. Delcour I, Spanoghe P, Uyttendaele M (2015) Literature review: impact of climate change on pesticide use. Food Res Int 68:7–15CrossRefGoogle Scholar
  7. Demissie AG, Kumar A (2014) Production and partial purification of laccase produced by Bacillus species isolated from contaminated soil. Int J Emerg Tech Adv Eng 4:17862–17869Google Scholar
  8. Dhiman K, Shirkot P (2015) Bioprospecting and molecular characterization of laccase producing bacteria from paper mills of himachal pradesh. Sect B Biol Sci 85:4Google Scholar
  9. Eduardo T, Bustos-Jaimes I, Le Sylvie B (2003) Potential use of oxidative enzymes for the detoxification of organic pollutants. Appl Catal B Environ 46:1–15CrossRefGoogle Scholar
  10. Garlapati D, Chandrasekaran M, Devanesan A, Mathimani T, Pugazhendhi A (2019) Role of cyanobacteria in agricultural and industrial sectors: an outlook on economically important byproducts. Appl Microbiol Biotechnol 103(12):4709–4721CrossRefGoogle Scholar
  11. Govarthanan M, Mythili R, Selvankumar T, Kamala-Kannan S, Rajasekar A, Chang Young-Cheol (2016) Bioremediation of heavy metals using an endophytic bacterium Paenibacillus sp. RM isolated from the roots of Tridax procumbens. 3 Biotech 6:242CrossRefGoogle Scholar
  12. Guo H, Lin C, Wang S, Jiang D, Zheng B, Liu Y, Qin W (2017) Characterization of a novel laccase producing Bacillus sp. A4 and its application in Miscanthus degradation. Bio Resour 12:4776–4794Google Scholar
  13. Jaya Mary J, Karthik C, Smita GR, Kumar S, Prabakar D, Kadirvelu K, Pugazhendhi A (2018) Biological approaches to tackle heavy metal pollution: a survey of literature. J Environ Manage 217:56–70CrossRefGoogle Scholar
  14. Jin XH, Heo PS, Hong JS, Kim NJ, Kim YY (2016) Supplementation of dried mealworm (Tenebrio molitor larva) on growth performance, nutrient digestibility and blood profiles in weaning pigs. Asian Aust J Anim Sci 29:979–986CrossRefGoogle Scholar
  15. Karimi K, Mohammad J, Taherzadeh A (2016) A critical review on analysis in pretreatment of lignocelluloses: degree of polymerization, adsorption/desorption, and accessibility. Bioresour Technol 230:348–356CrossRefGoogle Scholar
  16. Kim TH, Kim JS, Sunwoo C, Lee YY (2017) Pretreatment of corn stover by aqueous ammonia. Bioresour Technol 90:39–47CrossRefGoogle Scholar
  17. Kuddus M, Joseph B, Ramteke PW (2013) Bacterial laccase: recent update on production, properties and industrial applications. Biocatal Agric Biotechnol 2:333–338CrossRefGoogle Scholar
  18. Kumar A, Chauha BM (1993) Chemical composition and utilization of pearl millet sprouts. Mol Nutr Food Res 37(4):356–363Google Scholar
  19. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of Bacteriophage T4. Nature 227:680–685CrossRefGoogle Scholar
  20. Liu J, Tan L, Wang J, Wang Z, Ni H, Li L (2016) Complete biodegradation of chlorpyrifos by engineered Pseudomonas putida cells expressing surface immobilized laccases. Chemosphere 157:200–207CrossRefGoogle Scholar
  21. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the folin phenol reagent. J Biol Chem 193:265–275Google Scholar
  22. Lu R, Miyakoshi T (2012) Studies on acetone powder and purified rhus laccase immobilized on zirconium chloride for oxidation of phenols. Enzyme Res 8:375309Google Scholar
  23. Margot J, Granier CB, Maillard J, Blánquez P, Barry DA, Holliger C (2013) Bacterial versus fungal laccase: potential for micropollutant degradation. AMB Express 3:6CrossRefGoogle Scholar
  24. Mishra SK, Srivastava SK (2016) Production of extracellular laccase from bacterial strain Bacillus subtilis MTCC 1039 using different parameter. Biosci Biotech Res 13(3):1645–1650CrossRefGoogle Scholar
  25. Molina Guijarro JM, Pérez J, Muñoz Dorado J, Guillén F, Moya R, Hernández M, Arias ME (2009) Detoxification of azo dyes by a novel pH versatile, salt resistant laccase from Streptomyces ipomoea. Int Microbiol 12(1):13–21PubMedGoogle Scholar
  26. Narayanan PM, Jackson B, Raj AJ, Sevanan M (2015) Production of extracellular laccase from Bacillus subtilis MTCC 2414 using agro residues as a potential substrate. Biochem Res Int 1:9–18Google Scholar
  27. Pizzul L, Pilar Castillo M, Stenstcom J (2009) Degradation of glyphosate and other pesticides by ligninolytic enzymes. Biodegradation 20:751–759CrossRefGoogle Scholar
  28. Rodriguez Couto S, Toca Herrera JL (2006) Industrial and biotechnological applications of laccases: a review. Biotechnol Adv 24:500–513CrossRefGoogle Scholar
  29. Selvam K, Govarthanan M, Kamala-Kannan S, Govindharaju M, Senthilkumar B, Selvankumar T, Sengottaiyan A (2014) Process optimization of cellulase production from alkali-treated coffee pulp and pineapple waste using Acinetobacter sp. TSK-MASC. RSC Adv 4:13045CrossRefGoogle Scholar
  30. Selvam K, Selvankumar T, Radhika R, Srinivasan P, Sudhakar C, Senthilkumar B, Govarthanan M (2016a) Enhanced production of amylase from Bacillus sp. Using groundnut shell and cassava waste as a substrate under process optimization: waste to wealth approach. Biocatal Agric Biotechnol 7:250–256CrossRefGoogle Scholar
  31. Selvam K, Govarthanan M, Senthilkumar B, Senbagam D, Selvankumar T, Kamala-Kannan S, Sudhakar C (2016b) Optimization of protease production from surface modified coffee pulp waste and corncobs using Bacillus sp. by SSF. 3 Biotech 6:167Google Scholar
  32. Selvankumar T, Radhika R, Mythili R, Arunprakash S, Srinivasan P, Govarthanan M, Kim Hyunook (2017) Isolation, identification and characterization of arsenic transforming exogenous endophytic Citrobacter sp. RPT from roots of Pteris vittata. 3 Biotech 7:264CrossRefGoogle Scholar
  33. Sharma N, Kalra KL, Oberoi HS, Bansal S (2017) Optimization of fermentation parameters for production of ethanol from kinnow waste and banana peels by simultaneous saccharification and fermentation. Indian J Microbiol 47:310–316CrossRefGoogle Scholar
  34. Sharma V, Ayothiraman S, Dhakshinamoorthy V (2018) Production of highly thermo–tolerant laccase from novel thermophilic bacterium Bacillus sp. PC–3 and its application in functionalization of chitosan film. J Biosci Bioeng 127(6):672–678CrossRefGoogle Scholar
  35. Sonica S, Prince S, Nancy G, Prakram S, Chauhan P, Naveen G (2015) An extracellular thermo alkali stable laccase from Bacillus tequilensis SN4, with a potential to biobleach softwood pulp. 3 Biotech 5:175–185Google Scholar
  36. Upadhyay P, Shrivastava R, Agrawal PK (2016) Bioprospecting and biotechnological applications of fungal laccase. 3 Biotech 6:1–12CrossRefGoogle Scholar
  37. Wang Z, Denisi M, Vaida P, Chen B, Moshammerf K, Mohamed YS, Wang HS, Sioudh S, Rajih MA, Hooinghausi KK, Hansenf N, Dagautj P, Leone SR, Mani Sarathya S (2017) New insights into the low temperature oxidation of 2–methylhexane. Proc Combust Inst 36:373–382CrossRefGoogle Scholar
  38. Wolfenden RS, Wilson RL (1982) Radicalcations as reference chromogens in the kinetic studies of one–electron transfer reactions: pulse radiolysis studies of 2, 2′–azinobis–(3–ethylbenzthiazoline–6–sulphonate). J Chem Soc Perkin Trans 2:805–812CrossRefGoogle Scholar
  39. Zhao D, Zhang X, Cui D, Zhao M (2012) Characterization of a novel white laccase from the Deuteromycete fungus Myrothecium verrucaria NF05 and its decolourisation of dyes. PLoS ONE 7:38817CrossRefGoogle Scholar
  40. Zhou N, Li D, Wu S, He S, Chen Y, Bai Y, Zhou M, He J, Wang C (2015) Acceleration effect of sodium selenite on yeast growth and fermentative capability. J Gen Appl Microbiol 61:27–30CrossRefGoogle Scholar

Copyright information

© King Abdulaziz City for Science and Technology 2019

Authors and Affiliations

  1. 1.PG and Research Department of BiotechnologyMahendra Arts and Science College (Autonomous)Kalippatti, NamakkalIndia
  2. 2.Division of BiotechnologyCollege of Environmental and Bioresource Sciences, Chonbuk National UniversityIksanSouth Korea
  3. 3.Department of Medical MicrobiologyCollege of Health and Medical Sciences, Haramaya UniversityHararEthiopia

Personalised recommendations