Amylase Production Optimization and Textile Application of a Novel Bacillus Halotolerans SH1 Isolate from Textile Wastewater

Document Type : Original Article

Authors

1 Department of Textile Engineering, Yazd University, P.O. Box: 89195-741, Yazd, Iran

2 Department of Biology, Yazd University, P.O. Box: 89195-741, Yazd, Iran

Abstract

Textile wastewater usually contains toxic compound and a few bacteria survive in this condition. Microorganisms in textile wastewater can produce a variety of enzymes and biotechnology products. This study aimed to isolate bacteria that produce amylase and polyhydroxyalkanoates (PHAs) from textile waste. Both of these compounds are used in the textile industry. Amylase enzyme is used to enhance thread strength and PHAs for the packaging and production of suture thread, surgical meshes, cardiovascular fabrics, and conventional fibers. The highest enzyme activity was optimized using the response surface method (RSM). The isolate was identified as Bacillus halotolerans SH1 by determining the 16S rRNA gene sequence. Optimum conditions for maximum enzyme production (0.26 U/mg) were starch 5.5 g/L, temperature 37 °C, and incubation period 48 hours. The results showed that a PHA concentration of 0.27 g/L is obtained from this bacterium. The reduction in BOD, COD by bacteria was found to be 54.93 and 51.92 %, respectively. This study reports a new strain of Bacillus halotolerans SH1, isolated from toxic textile wastewater, which is capable of producing both PHA and α-amylase enzyme, while also significantly reducing BOD and COD levels, showing its multifunctional potential in environmental and industrial applications, especially in the textile industry. 

Keywords

Main Subjects

References

  1. Cha´vez-Camarillo GMa, Lopez-Nuñez PV, Jime´nez- Nava1 RA, Aranda-Garcıa E, Cristiani-Urbina E. Production of extracellular α-amylase by single-stage steady-state continuous cultures of Candida wangnamkhiaoensis in an airlift bioreactor. PLOS ONE. 2022; 17(3): e0264734. https://doi.org/ 10.1371/ journal.pone.0264734.
  2. Tanyildizi MS, Ozer D, Elibol M. Optimization of a-amylase production by Bacillus sp. using response surface methodology. Process Biochem. 2005; 40(7): 2291-2296. https://doi.org/10.1016/j.procbio.2004.06. 018.
  3. Kumari N, Sushil, Rani B, Malik K, Avtar R. Microbial amylases: An overview on recent advancement. J Entomol Zool Stud. 2019; 7(1): 198-205. 
  4. Msarah MJ, Ibrahim I, Hamid AA, Aqma WS. Optimisation and production of alpha amylase from thermophilic Bacillus spp. and its application in food waste biodegradation. Heliyon. 2020; 6(6): e04183. https://doi.org/10.1016/j.heliyon.2020.e04183.
  5. Andualem B. Isolation and screening of amylase producing thermophilic spore forming Bacilli from starch rich soil and characterization of their amylase activities using submerged fermentation. Int Food Res J. 2014; 21(2): 831-837.
  6. Hallol M, Helmy O, Shawky AE, El-Batal A, Ramadan M. Optimization of alpha-amylase production by a local bacillus paramycoides isolate and immobilization on chitosan-loaded barium ferrite nanoparticles. Fermentation. 2022; 8(5):241. https://doi.org/10.3390/fermentation8050241.
  7. Mojsov K. Application of enzymes in the textile industry. In: II International Congress “Engineering, Ecology and Materials in the Processing Industry”; 2011; Jahorina. p. 230-239.
  8. Sundarram A, Krishna Murthy ThP. α-Amylase production and applications: a review. J Appl Environ Microbiol. 2014; 2(4):166-175. https://doi.org/10. 12691/ jaem-2-4-10.
  9. Rehman A, Saeed A, Asad W, Khan I, Hayat A, Rehman MU, Shah TA, Sitotaw B, Dawoud T, Bourhia M. Eco‑friendly textile desizing with indigenously produced amylase from Bacillus cereus AS2. Sci Report. 2023; 13(1). https://doi.org/10. 1038/s41598-023-38956-3.
  10. Farooq MA, Ali Sh, Hassan A, Tahir HM, Mumtaz S, Mumtaz Sh.  Biosynthesis and industrial applications of α‑amylase: a review. Arch Microbiol. 2021; 203(4): 1281-1292. https://doi.org/10.1007/s00203-020-02128-y.
  11. Mostafa FA, Wehaidy HR, El‑Hennawi HM, Mahmoud SA,  Sharaf  S, Saleh SAA. Statistical optimization of α-amylase production from novel local isolated bacillus spp. NRC1 and its textile applications. Catal  Lett. 2024; 154:3264-3275. https://doi.org/10.1007/s10562-023-04545-2.
  12. Far BE, Ahmadi Y, Khosroushahi AY, Dilmaghani A. Microbial alpha-amylase production: progress, challenges and perspectives. Adv Pharm Bull. 2020; 10(3):350-358. https://doi.org/10.34172/apb.2020.043.
  13. Adrio JL, Demain AL. Microbial Enzymes: Tools for biotechnological processes. biomolecules. 2014; 4(1):117-139. https://doi.org/10.3390/biom4010117.
  14. Konsoula Z, Liakopoulou-Kyriakides M. Co-production of alpha-amylase and beta-galactosidase by Bacillus subtilis in complex organic substrates. Bioresour Technol. 2007;98(1):150-157. https://doi. org/ 10.1016/j.biortech.2005.11.001.
  15. Gamal RF, Abou-Taleb KhA, Abd-Elhalim BT. Isolation, identification and production of amylases from thermophilic spore forming bacilli using starch raw materials under submerged culture. AASCIT J Biosci. 2017; 3(6): 52-68.
  16. Sonunea N, Garode A. Isolation, characterization and identification of extracellular enzyme producer Bacillus licheniformis from municipal wastewater and evaluation of their biodegradability. Biotechnol Res Innov. 2018;2(1):37-44. https://doi.org/10.1016/j. biori. 2018.03.001.
  17. Olaiya NG, Oyekanmi AA, Hanafiah MM, Olugbade TO, Adeyeri MK, Olaiya FG. Enzyme-assisted extraction of nanocellulose from textile waste: A review on production technique and applications. Bioresour Technol Rep. 2022;19:101183. https://doi. org/ 10.1016/j.biteb.2022.101183.
  18. Shukla A, Parmar P, Goswami D, Gehlot Y, Vala J, Parmar  N, Saraf M. Microbial technologies in textile industries: an elixir for the greener environment. In: Green Chemistry for Sustainable Textiles: Modern Design and Approaches. Woodhead Publishing; 2021. p. 173-189.
  19. Cavaliere Ch, Capriotti AL, Cerrato A. Identification and quantification of polycyclic aromatic hydrocarbons in polyhydroxyalkanoates produced from mixed microbial cultures and municipal organic wastes at pilot scale. Molecules. 2021; 26(3):539. https://doi.org/10.3390/molecules26030539.
  20. Nielsen Ch. Investigations of polyhydroxyalkanoate secretion and production using sustainable carbon sources [master’s thesis], Biological Engineering, Utah State Univ.; 2018.
  21. Cavaliere Ch, Capriotti AL, Cerrato A, Lorini  L, Montone  CM, Valentino F, et al. Identification and quantification of polycyclic aromatic hydrocarbons in polyhydroxyalkanoates produced from mixed microbial cultures and municipal organic wastes at pilot scale. Molecules. 2021; 26(3):539. https://doi.org/10.3390/molecu les26030539 
  22. Shahi Z, Khajeh Mehrizi M, Mirbagheri Firoozabad MS. Introduction of Polyhydroxyalkanoates (PHAs) and Their Application in the Textile Industry. In: Biopolymers in the Textile Industry: Springer Nature Singapore, 2024. p. 29-40.
  23. Bezerraa MA, Santelli RE, Oliveira EP, Villar LS, Escaleira LA. Response surface methodology (RSM) as a tool for optimization in analytical chemistry. Talanta. 2008; 76:965-977. https://doi.org/10.1016/j. talanta.2008.05.019.
  24. Abdel-Fattah YR, Soliman NA, El-Toukhy NM, El-Gendi H, Ahmed RS. Production, purification, and characterization of thermostable α-amylase produced by Bacillus licheniformis Isolate AI20. J Chem. 2013; 2. https://doi.org/10.1155/2013/673173.
  25. Krishnamoorthy K. Landge AV. Statistical optimisation of fermentation media for novel clostridium novyi-nt using response surface methodology. Braz Arch Biol Technol. 2018;61:e181 80026. https://doi.org/10.1590/1678-4324-20181800 26.
  26. Lau HL, Wong FWF, Abd Rahman RNZR, Mohamed MSh, Ariff AB, Hii SL. Optimization of fermentation medium components by response surface methodology (RSM) and artificial neural network hybrid with genetic algorithm (ANN-GA) for lipase production by Burkholderia cenocepacia ST8 using used automotive engine oil as substrate. Biocatal Agric Biotechnol. 2023; 50:102696. https://doi.org/ 10.1016/j.bcab.2023.102696.
  27. Sharif S, Shah AH, Fariq A, Jannat S, Rasheed S, Yasmin A. Optimization of amylase production using response surface methodology from newly isolated thermophilic bacteria. Heliyon. 2023; 9(1):e12901. https://doi.org/10.1016/j.heliyon.2023.e12901.
  28. Al-Bedak OAM, Moharram AM,  Ameen F, Stephenson SL,  Idres MMM, Yasser MM. Optimizing conditions for augmented production of amylase by Talaromyces islandicus AUMC 11391. Electronic J Biotechnol. 2025;75:39-48. https://doi. org/ 10.1016/j.ejbt.2025.01.008.
  29. Liu Z, Smith1 SR. Enzyme recovery from biological wastewater treatment. Waste Biomass Valorization. 2021; 12:4185-4211. https://doi.org/10.1007/s12649-020-01251-7.
  30. Ajmal AW, Saroosh S, Mulk Sh. Bacteria isolated from wastewater irrigated agricultural soils adapt to heavy metal toxicity while maintaining their plant growth promoting traits. Sustainability. 2021;13(14): 7792. https://doi.org/10.3390/su13147792.
  31. Kurahman OT, Yuliawati A, Haerunnisa L, Supriyatna A, Cahyanto T, Suryani Y, et al. The isolation and identification bacteria on jallalah animal (study on the feeding tilapia (oreochromis niloticus) with chicken manure as foods). Elkawnie: J Islamic Sci Technol. 2023;6(2):222. https://doi.org/10.22373/ ekw.v6i2.7770.
  32. Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Struh K. Short protocols in molecular biology, John Wiley & Sons, New York, NY, USA. 1999.
  33. Cihan AC, Tekin N, Ozcan B, Cokmus C. The genetic diversity of genus Bacillus and the related genera revealed by 16S rRNA gene sequences and ardra analysis isolated from geothermal regions of Turkey. Brazilian J Microbiol. 2012; 43(1):309-324. https://doi.org/10.1590/S1517-838220120001000037.
  34. Poorani E, Saseetharan M, Dhevagi P. Lasparaginase production and molecular identification of marine Streptomyces sp. strain EPD 27. Int J Integr Biol. 2009; 7(3):150-155.
  35. Padhiar AR, Kommu S. Isolation, Characterization and Optimization of Bacteria producing Amylase. Inter J Adv Res Biol Sci. 2016; 3(7):1-7. 
  36. Boshagh F. Measurement methods of carbohydrates in dark fermentative hydrogen production- A review. Int J Hydrogen Energy. 2021; 46(47):24028-24050. https://doi.org/10.1016/j.ijhydene.2021.04.204.
  37. Mageswari A, Subramanian P, Chandrasekaran S, Sivashanmugam K, Babu S, Gothandam KM. Optimization and immobilization of amylase obtained from halotolerant bacteria isolated from solar salterns. J Genet Eng Biotechnol. 2012; 10(2): 201-208. https://doi.org/10.1016/j.jgeb.2012.09.001.
  38. Bernal-Ruiz M, Correa-Lozano A, Gomez-Sánchez L, Quevedo-Hidalgo B, Rojas-Pérez LC, García-Castillo C, et al.. Brewer’s spent grain as substrate for enzyme and reducing sugar production using Penicillium sp. HC1. Revista de la Academia Colombiana de Ciencias Exactas, Físicas y Naturales. 2021; 45(176): 850-863. https://doi.org/10.18257/ raccefyn. 1379.
  39. Krowiak AW, Chojnacka K, Podstawczyk D, Dawiec A, Bubała K. Application of response surface methodology and artificial neural network methods in modelling and optimization of biosorption process. Bioresour Technol. 2014; 160:150-160. https://doi. org/ 10.1016/j.biortech.2014.01.021.
  40. Haji A, Mousavi Shoushtari A, Mazaheri F, Tabatabaeyan SE. RSM optimized self-cleaning nano-finishing on polyester/wool fabric pretreated with oxygen plasma. J Text Inst. 2016; 107(8): 985-994. https://doi.org/10.1080/00405000.2015.1077023.
  41. Hadizadeh M, Khajeh Mehrizi M, Shahi Z. Application of response surface methodology for modeling the color strength of natural hair colorant. J Text Polym. 2019; 7(1): 15-23.
  42. Bhattacharya S. Central composite design for response surface methodology and its application in pharmacy. In: Response surface methodology in engineering science. IntechOpen, 2021.‏
  43. Lageveen RG, Huisman GW, Preusting H, Ketelaar P, Eggink G, Witholt B. Formation of polyesters by Pseudomonas eleovans: Effect of substrates on formation and composition of poly-(R)-3- hydroxyl-alkanoates and poly-(R)-3-hydroxyalkanoates. Appl Environ Microbiol. 1988; 54(12):2924-2932. https://doi.org/10.1128/aem.54.12.2924-2932.1988.
  44. Mascarenhas J, Aruna K. Screening of polyhydroxy-alkonates (PHA) accumulating bacteria from diverse habitats. J Glob Biosci. 2017;  6(3):4835-4848. https://doi.org/10.13140/RG.2.2.29966.72005.
  45. Clesceri LS, Greenberg AE, Eaton AD. American Public Health Association (APHA); American Water Works Association (AWWA); Water Environment Federation (WEF). Standard Methods for the Examination of Water and Wastewater, 1998, 2671.‏
  46. Karyab  H, Hamidi Z, Ghotbinia F, Mosa Khani Z, Nazeri N. Application of the central composite design and response surface methodology for optimization of reactive color removal from aqueous solutions using dicyandiamide-formaldehyde resin modified by ammonium chloride. Desalin Water Treat. 2023; 308:217-228. https://doi.org/10.5004/dwt.2023.29829.
  47. Fajdek-Bieda A, Perec A, Radomska-Zalas A. Application of RSM Method for Optimization of Geraniol Transformation Process in the Presence of Garnet. Int J Mol Sci. 2023; 24(3):2689; https://doi.org/10.3390/ijms24032689.
  48. Adetiloye OA, Solomon BO, Omolaiye JA, Betiku E. Optimization of thermostable amylolytic enzyme production from Bacillus cereus isolated from a recreational warm spring via Box Behnken design and response surface methodology. Microb Cell Fact. 2025; 24:87. https://doi.org/10.1186/s12934-025-02709-w.
  49. Jia YY, Zhang L. A Process for preparing polyethylene wax microspheres and optimization of their dissolution precipitation by response surface methodology. J Appl Polym Sci. 2013; 129(3):1476-1483. https://doi.org/10.1002/app.38847.
  50. Niyonzima FN. Production of microbial industrial enzymes. acta scientific microbiology. 2019; 2(12):75-89. https://doi.org/10.31080/ASMI.2019.02.0434. 
  51. 51.Wang X, Xia K, Yang X, Tang Ch. Growth strategy of microbes on mixed carbon sources. Nat Commun. 2019; 10(1): 1279. https://doi.org/10. 1038/s41467-019-09261-3.
  52. Mengesha Y, Tebeje A, Tilahun B. A review on factors influencing the fermentation process of teff (Eragrostis teff) and other cereal-based ethiopian injera. Int J Food Sci. 2022. https://doi.org/10. 1155/ 2022/4419955.
  53. Rakkan Th, Chana N, Sangkharak K. The integration of textile wastewater treatment with polyhydroxy-alkanoate production using newly isolated Enterobacter Strain TS3. Waste Biomass Valor. 2022; 13: 571-582. https://doi.org/10.1 007/ s12649-021-01504-z.
  54. Shahi Z, Khajeh Mehrizi M, Mirbagheri Firoozabad MS. Decolorization of Textile Dye by Isolated Bacterial Strains from Textile Waste and Their Ability to Produce Polyhydroxyalkanoate. Prog. Color Colorant Coat. 2025; 18:99-111. https://doi.org/10. 30509/PCCC.2024.167299.1292.
Progress in Color, Colorants and Coatings
Volume 19, Issue 1 - Serial Number 60
September 2025
Pages 83-95

Files

Share

How to cite

Statistics