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Glutamic Acid Independent Production of Bioflocculants by Bacillus subtilis UPMB13

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Abstract

Bacillus subtilis UPMB13 was found to be an L-glutamic acid independent producer of extracellular polymeric substances (EPS) with bioflocculation properties. Optimum production of the bioflocculant was found to be at the early stage of cell propagation of 24–72 h of fermentation. At a limited nutrient input of 100 mL tryptic soy broth, the flocculating activities were found to be negatively correlated (p < 0.01) with growth as it continued to decline after 72 h, while cell growth proliferated further. Ample nutrient supply may prolong bioflocculant production with flocculating activities of 90 % and higher, while excess oxygen supply may promote rapid growth that can lead to poor flocculation due to the re-use of the bioflocculant as a substitute for food during starvation. Bioflocculant production occurred at best at 25-30 °C incubation temperature and at the initial pH medium of 7 to 8. The bioflocculant was proven to be extracellularly produced as the broth and the supernatant possessed the ability to flocculate the suspended kaolin particles. Bioflocculant productions by UPMB13 were hereditarily stable among succeeding progenies, hence, proving genetic competency. About 0.90 g of purified bioflocculant were collected from 1 L culture broth of UPMB13 under the optimized fermentation conditions.

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References

  • Aljuboori AHR, Idris A, Abdullah N, Mohamad R (2013) Production and characterization of a bioflocculant produced by Aspergillus flavus. Bioresource Technol 127:489–493. doi:10.1016/j.biortech.2012.09.016

    Article  Google Scholar 

  • Amir HG, Shamsuddin ZH, Halimi MS, Ramlan MF, Marziah M (2003) N2 fixation, nutrient accumulation and plant growth promotion by rhizobacteria in association with oil palm seedlings. Pakistan J Biol Sci 6:1269–1272

    Article  Google Scholar 

  • Bajaj I, Singhal R (2011a) Poly (glutamic acid): an emerging biopolymer of commercial interest. Bioresource Technol 102:5551–5561. doi:10.1016/j.biortech.2011.02.047

    Article  Google Scholar 

  • Bajaj I, Singhal R (2011b) Flocculation properties of poly(γ-glutamic acid) produced from Bacillus subtilis isolate. Food Bioprocess Tech 4:745–752. doi:10.1007/s11947-009-0186-y

    Article  Google Scholar 

  • Bhunia B, Mukhopadhy D, Goswami S, Mandal T, Dey A (2012) Improved production, characterization and flocculation properties of poly (γ)-glutamic acid produced from Bacillus subtilis. J Biochem Technol 3:389–394

    Google Scholar 

  • Chen X, Chen S, Sun M, Yu Z (2005) Medium optimization by response surface methodology for poly-γ-glutamic acid production using dairy manure as the basis of a solid substrate. Appl Microbiol Biot 69:390–396. doi:10.1007/s00253-005-1989-z

    Article  Google Scholar 

  • Cosa S, Ugbenyen AM, Mabinya LV, Rumbold K, Okoh AI (2013) Characterization and flocculation efficiency of a bioflocculant produced by a marine Halobacillus. Environ Technol 34:2671–2679. doi:10.1080/09593330.2013.786104

    Article  Google Scholar 

  • Devesa-Rey R, Bustos G, Cruz J, Moldes A (2012) Evaluation of non-conventional coagulants to remove turbidity from water. Water Air Soil Poll 223:591–598. doi:10.1007/s11270-011-0884-8

    Article  Google Scholar 

  • Dierksen KP, Sandine WE, Trempy JE (1997) Expression of ropy and mucoid phenotypes in Lactococcus lactis. J Dairy Sci 80:1528–1536. doi:10.3168/jds.S0022-0302(97)76082-X

    Article  Google Scholar 

  • Ito Y, Tanaka T, Ohmachi T, Asada Y (1996) Glutamic acid independent production of poly (y-glutamic acid) by Bacillus subtilis TAM-4. Biosci Biotech Bioch 60:1239–1242. doi:10.1271/bbb.60.1239

    Article  Google Scholar 

  • Kimura K, Tran LSP, Uchida I, Itoh Y (2004) Characterization of Bacillus subtilis gamma-glutamyltransferase and its involvement in the degradation of capsule poly-gamma-glutamate. Microbiol 150:4115–4123. doi:10.1099/mic.0.27467-0

    Article  Google Scholar 

  • Korsten L, Cook N (1996) Optimizing culturing conditions for Bacillus subtilis. South Afric Avocado Grow Assoc Yearbook 19:54–58

    Google Scholar 

  • Liu LF, Cheng W (2010) Characteristics and culture conditions of a bioflocculant produced by Penicillium sp. Biomed Environ Sci 23:213–218. doi:10.1016/S0895-3988(10)60055-4

    Article  Google Scholar 

  • Mahmoud DAR (2006) Isolation of polyglutamic acid flocculant producing bacteria from extreme egyptian environments. Journal of Applied Science Research 2:608–612

    Google Scholar 

  • Maximova N, Dahl O (2006) Environmental implications of aggregation phenomena: current understanding. Curr Opin Colloid In 11:246–266. doi:10.1016/j.cocis.2006.06.001

    Article  Google Scholar 

  • Muthulakshmi L, Nellaiah H, Busi S (2013) Production and characterization of a novel bioflocculant from Klebsiella sp. Curr Biot 2:53–58

    Article  Google Scholar 

  • Ntsaluba L, Nwodo UU, Mabinya L, Okoh A (2013) Studies on bioflocculant production by a mixed culture of Methylobacterium sp. obi and Actinobacterium sp. mayor. BMC Biotechnol 13:1–7. doi:10.1186/1472-6750-13-62

    Article  Google Scholar 

  • Nwodo UU, Green E, Mabinya LV, Okaiyeto K, Rumbold K, Obi LC, Okoh AI (2014) Bioflocculant production by a consortium of Streptomyces and Cellulomonas species and media optimization via surface response model. Colloid Surface B 116:257–264. doi:10.1016/j.colsurfb.2014.01.008

    Article  Google Scholar 

  • Nwodo UU, Okoh AI (2012) Characterization and flocculation properties of biopolymeric flocculant (glycosaminoglycan) produced by Cellulomonas sp. Okoh J Appl Microbiol 114:1325–1337. doi:10.1111/jam.12095

    Article  Google Scholar 

  • Okaiyeto K, Nwodo UU, Mabinya LV, Okoli AS, Okoh AI (2015) Characterization of a bioflocculant (MBF-UFH) produced by Bacillus sp. AEMREG7. Int J Mol Sci 16:12986–13003. doi:10.3390/ijms160612986

    Article  Google Scholar 

  • Patil S, Salunkhe R, Patil C, Patil D, Salunke B (2010) Bioflocculant exopolysaccharide production by Azotobacter indicus using flower extract of Madhuca latifolia L. Appl Biochem Biotech 162:1095–1108. doi:10.1007/s12010-009-8820-8

    Article  Google Scholar 

  • Patil SV, Bathe GA, Patil AV, Patil RH, Salunkea BK (2009) Production of bioflocculant exopolysaccharide by Bacillus subtilis. Adv Biot 8:14–17

    Google Scholar 

  • Peirui L, Zongwei L, Zongyi L, Guangyong Q, Yuping H (2008) Screening of bioflocculant-producing strain by ion implantation and flocculating characteristics of bioflocculants. Plasma Sci Technol 10:394. doi:10.1088/1009-0630/10/3/26

    Article  Google Scholar 

  • Prasertsan P, Wichienchot S, Doelle H, Kennedy JF (2008) Optimization for biopolymer production by Enterobacter cloacae WD7. Carbohyd Polym 71:468–475. doi:10.1016/j.carbpol.2007.06.017

    Article  Google Scholar 

  • Richard A, Margaritis A (2003) Rheology, oxygen transfer, and molecular weight characteristics of poly(glutamic acid) fermentation by Bacillus subtilis. Biotechnol Bioeng 82:299–305. doi:10.1002/bit.10568

    Article  Google Scholar 

  • Salehizadeh H, Yan N (2014) Recent advances in extracellular biopolymer flocculants. Biotechnol Adv 32:1506–1522. doi:10.1016/j.biotechadv.2014.10.004

    Article  Google Scholar 

  • Shih IL, Van YT (2001) The production of poly-(γ-glutamic acid) from microorganisms and its various applications. Bioresource Technol 79:207–225. doi:10.1016/S0960-8524(01)00074-8

    Article  Google Scholar 

  • Shih L, Wu JY (2009) Biosynthesis and application of poly (γ-glutamic acid). In: Bernd R (ed) Microbial production of biopolymers and polymer precursors: applications and perspectives. Horizon Scientific Press, Norfolk, pp. 101–141

    Google Scholar 

  • Shih IL, Wu PJ, Shieh CJ (2005) Microbial production of a poly(γ-glutamic acid) derivative by Bacillus subtilis. Process Biochem 40:2827–2832. doi:10.1016/j.procbio.2004.12.009

    Article  Google Scholar 

  • Su X, Shen X, Ding L, Yokota A (2012) Study on the flocculability of the Arthrobacter sp., an actinomycete resuscitated from the VBNC state. World J Microb Biot 28:91–97. doi:10.1007/s11274-011-0795-2

    Article  Google Scholar 

  • Subramanian SB, Yan S, Tyagi RD, Surampalli RY (2009) Bioflocculants. In: Tyagi RD, Surampalli RY, Yan S, Zhang TC, Kao CM, Lohani BN (eds) Sustainable sludge management. American Society of Civil Engineers, Reston, pp. 146–167

    Chapter  Google Scholar 

  • Wang X, Zhang Y, Zhong W (2008) Poly-γ-glutamic acid production by novel isolated Bacillus subtilis zjutzy and its flocculating character. J Biotechnol 136:S43

    Article  Google Scholar 

  • Wei W, Fang M, Xiuli Y, Aijie W (2008) Purification and characterization of compound bioflocculant. In: The 2nd international conference on bioinformatics and biomedical engineering. Shanghai, IEEE, pp. 1127–1130. doi:10.1109/ICBBE.2008.275

    Google Scholar 

  • Wong YS, Ong SA, Teng TT, Aminah LN, Kumaran K (2012) Production of bioflocculant by Staphylococcus cohnii sp. from palm oil mill effluent (POME). Water Air Soil Poll 223:3775–3781. doi:10.1007/s11270-012-1147-z

    Article  Google Scholar 

  • Wu JY, Ye HF (2007) Characterization and flocculating properties of an extracellular biopolymer produced from a Bacillus subtilis DYU1 isolate. Process Biochem 42:1114–1123. doi:10.1016/j.procbio.2007.05.006

    Article  Google Scholar 

  • Xu H, Jiang M, Li H, Lu D, Ouyang P (2005) Efficient production of poly(γ-glutamic acid) by newly isolated Bacillus subtilis NX-2. Process Biochem 40:519–523. doi:10.1016/j.procbio.2003.09.025

    Article  Google Scholar 

  • Yang X (2011) Preparation and characterization of γ-poly (glutamic acid) copolymer with glycol diglycidyl ether. Procedia Environmental Sciences 8:11–15. doi:10.1016/j.proenv.2011.10.004

    Article  Google Scholar 

  • Yokoi H, Arima T, Hirose J, Hayashi S, Takasaki Y (1996) Flocculation properties of poly([γ]-glutamic acid) produced by Bacillus subtilis. J Ferment Bioeng 82:84–87. doi:10.1016/0922-338X(96)89461-X

    Article  Google Scholar 

  • Yokoi H, Natsuda O, Hirose J, Hayashi S, Takasaki Y (1995) Characteristics of a biopolymer flocculant produced by Bacillus sp. PY-90. J Ferment Bioeng 79:378–380. doi:10.1016/0922-338X(95)94000-H

    Article  Google Scholar 

  • Yu W, Chen Z, Shen L, Wang Y, Li Q, Yan S, Zhong CJ, He N (2015) Proteomic profiling of Bacillus licheniformis reveals a stress response mechanism in the synthesis of extracellular polymeric flocculants. Biotechnol Bioeng 9999:1–10. doi:10.1002/bit.25838

    Google Scholar 

  • Zhang H, Zhu J, Zhu X, Cai J, Zhang A, Hong Y, Huang J, Huang L, Xu Z (2012) High-level exogenous glutamic acid-independent production of poly-(γ-glutamic acid) with organic acid addition in a new isolated Bacillus subtilis C10. Bioresource Technol 116:241–246. doi:10.1016/j.biortech.2011.11.085

    Article  Google Scholar 

  • Zulkeflee Z, Aris AZ, Shamsuddin ZH, Yusoff MK (2012) Cation dependence, pH tolerance, and dosage requirement of a bioflocculant produced by Bacillus spp. UPMB13: flocculation performance optimization through kaolin assays. Sci World J. doi:10.1100/2012/495659

    Google Scholar 

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Acknowledgments

This research has been funded by the Spanish Ministerio de Economia y Competitividad (project CTM2015-69513-R). Dimitrios Komilis thanks Tecniospring for the financial support.

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Authors and Affiliations

  1. Department of Environmental Sciences, Faculty of Environmental Studies, Universiti Putra Malaysia, 43400, UPM Serdang, Selangor, Malaysia

    Zufarzaana Zulkeflee

  2. Composting Research Group, Department of Chemical Engineering, Escola d’Enginyeria, Universitat Autònoma de Barcelona, 08913-Bellaterra, Cerdanyola del Vallès, Barcelona, Spain

    Zufarzaana Zulkeflee, Dimitrios Komilis & Antoni Sánchez

  3. Department of Land Management, Faculty of Agriculture, Universiti Putra Malaysia, 43400, UPM Serdang, Selangor, Malaysia

    Zulkifli H. Shamsuddin

  4. Environmental Forensics Research Centre, Faculty of Environmental Studies, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia

    Ahmad Zaharin Aris

  5. UPM Consultancy and Services Sdn. Bhd., Universiti Putra Malaysia, 43400, UPM Serdang, Selangor, Malaysia

    Mohd Kamil Yusoff

Authors
  1. Zufarzaana Zulkeflee
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  2. Zulkifli H. Shamsuddin
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  3. Ahmad Zaharin Aris
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  4. Mohd Kamil Yusoff
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  5. Dimitrios Komilis
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  6. Antoni Sánchez
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Correspondence to Dimitrios Komilis.

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Zulkeflee, Z., Shamsuddin, Z.H., Aris, A.Z. et al. Glutamic Acid Independent Production of Bioflocculants by Bacillus subtilis UPMB13. Environ. Process. 3, 353–367 (2016). https://doi.org/10.1007/s40710-016-0161-3

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  • DOI: https://doi.org/10.1007/s40710-016-0161-3

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