Abstract
Over a hundred of halophilic/halotolerant microorganisms were screened for alkaline protease production. The bacterium showing the highest enzyme production was characterized and identified as Bacillus halodurans US193 on the basis of 16S rRNA gene analysis. It was alkalophilic, thermophilic and halotolerant since it grew optimally at pH 9.7 and 50 °C with tolerance of up to 125 g NaCl l−1. The alkaline protease was purified 4.9 times with about 40186.1 U/mg as specific activity. It exhibited optimal activity at pH 10, 70 °C and 0.25 M NaCl with perfect stability at wide ranges of pH (6–12), temperatures (30–60 °C) and NaCl concentrations (0–2 M). The serine alkaline protease maintained high stability in the presence of Cu2+, Mg2+, Ba2+ and Ca2+ ions, various organic solvents [50% (v/v)] and ionic and non ionic detergent additives. In addition, it was more compatible with various commercialized detergents than other reported detergent proteases, and was very efficient in blood stain removal. These findings let B. halodurans US193 alkaline protease be an ideal candidate for many industrial processes at harsh conditions, especially as a bio-additive in detergent industry.
Similar content being viewed by others
References
Ben Ali M, Mhiri S, Mezghani M, Bejar S (2001) Enzyme Microb Technol 28:537
Ben Ali M, Khemakhem B, Robert, Haser R, Bejar S (2006) Biochem J 394:51
Kirk O, Borchert TV, Fuglsang CC (2002) Curr Opin Biotech 13(3):45
Banik RM, Prakash M (2004) Microbiol Res 159:135
Genckal H, Tarib C (2006) Enzyme Microb Tech 39:703
Niyonzima FN, More S (2015) Prep Biochem Biotechnol 45:233
Jain D, Pancha I, Mishra SK, Shrivastav A, Mishra S (2012) Bioresource Technol 115:228
Sinha R, Srivastava AK, Khare SK (2014) Prep Biochem Biotechnol 44:680
Santos AFD, Pacheco CA, Valle RD, Seldin L, Santos ALD (2014) Appl Biochem Biotechnol 17:2748
Selim S, Hagagy N, Abdel Aziz M, El-Meleigy S, Pessione E (2014) Nat Prod Res 28:1476
Awad HM, Mostafa el-SE, Saad MM, Selim HM, Hassan HM (2013) Indian J Biochem Biophys 50:305
Das Sarma S, Arora P (2002) In: Nature Publishing Group (ed) Encyclopedia of life sciences, Nature Publishing Group, London, pp 458–466
Oren A (2006) Prokaryotes 2:263
Komagata K (1985) In: Hasegawa T (ed) Classification and identification of microorganisms. Japan Scientific Societies, Tokyo, pp 99–160
Tamura K, Stecher G, Peterson D, Filipski A, Kumar S (2013) Mol Biol Evol 30:2725
Kembhavi AA, Buttle DJ, Knight CG, Barret AJ (1993) Arch Biochem Biophys 303:208
Lin YC (1969) J Biol Chem 244:789
Bradford MM (1976) A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principal of protein–dye binding. Anal Biochem 72:248–254
Hmidet N, El-Hadj Ali N, Haddar A, Kanoun S, Sellami-Kamoun A, Nasri M (2009) Biochem Eng J 47:71
Claus D, Berkeley CW (1986) In: Sneath PHA (ed) Bergey’s manual of systematic bacteriology, vol 2. Williams, Wilkins, Baltimore, pp 1105–1139
Kitada M, Hashimoto M, Kudo T, Horikoshi K (1994) J Bacterial 176:6464
Takami H, Horikoshi K (1999) Biosci Biotechnol Biochem 63:943
Ito M, Xu H, Guffanti AA, Wei Y, Zvi L, Clapham DE et al (2004) Proc Natl Acad Sci USA 101:10566
Janto B, Ahmed A, Ito M, Liu J, Hicks DB, Pagni S et al (2011) Environ Microbiol 13(12):3289
Yin L, Xue Y, Ma Y (2015) PLoS One 10(6):e0128649. doi:10.1371/journal.pone.0128649
Shivanand P, Jayaraman G (2009) Process Biochem 44:1088
Sellami-Kamoun A, Haddar A, El-Hadj Ali N, Ghorbel-Frikha B, Kanoun S, Nasri M (2008) Microbiol Res 163:299
Haddar A, Agrebi R, Bougatef A, Hmidet N, Sellami-Kamoun A, Nasri M (2009) Bioresour Technol 100:3366
Lee S, Jang DJ (2001) Biophys J 81:2972
Annamalai N, Rajeswari MV, Vijayalakshmi S, Balasubramanian T (2013) Bioprocess Biosyst Eng 36:873
Shrinivas D, Naik GR (2011) Int Biodeter Biodegr 65:29
Jaouadi B, Ellouz-Chaabouni S, Ben Ali M, Ben Messaoud E, Naili B, Dhouib A, Bejar S (2009) Biotechnol Bioprocess Eng 14:503
Ben elhoul M, Zarai Jaoudi N, Rekik H, Bejar W, Boulkour Touioui S, Hmidi M, Badis A, Bejar S, Jaouadi B (2015) Int J Biol Macromol 79:871
Benkiar A, Zaraî Jaouadi N, Badis A, Rebzani F, Boulkour Touioui S, Rekik H, Naili B, Ferradji FZ, Bejar S, Jaouadi B (2013) Int Biodeter Biodegr 83:129
Raval VH, Pillai S, Rawal CM, Singh SP (2014) Process Biochem 49:955
Venugopal M, Saramma AV (2006) Process Biochem 41:1239
Bouacem K, Bouanane-Darenfed A, Ben Elhoul M, Hmida-Sayari A, Hacene H, Ollivier B, Fardeau ML, Jaouadi B, Bejar S (2015) Int J Biol Macromol 81:299
Acknowledgements
This work was funded by the Tunisian Ministry of Higher Education and Scientific Research and Technology (contract program LMBEE-CBS, Grant No. LR15CBS06).
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
No conflict of interest declared.
Rights and permissions
About this article
Cite this article
Daoud, L., Hmani, H., Ben Ali, M. et al. An Original Halo-Alkaline Protease from Bacillus halodurans Strain US193: Biochemical Characterization and Potential Use as Bio-Additive in Detergents. J Polym Environ 26, 23–32 (2018). https://doi.org/10.1007/s10924-016-0916-y
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10924-016-0916-y