African Journal of Microbiology Research
Subscribe to AJMR
Full Name*
Email Address*

Article Number - 512797E63282


Vol.11(11), pp. 440-449 , March 2017
DOI: 10.5897/AJMR2016-8379
ISSN: 1996-0808



Full Length Research Paper

Enhanced production of alkaline protease by Aspergillus niger DEF 1 isolated from dairy form effluent and determination of its fibrinolytic ability



Suseela Lanka
  • Suseela Lanka
  • Department of Biotechnology, Krishna University, Machilipatnam, 521001, Andhra Pradesh, India
  • Google Scholar
CH. Anjali
  • CH. Anjali
  • Department of Biochemistry, Krishna University Dr. MRAR PG Centre, Nuzvid, 521 201, Andhra Pradesh, India
  • Google Scholar
Muralidhar Pydipalli
  • Muralidhar Pydipalli
  • Vimta Labs, Cherlapally, Hyderabad, 500 007, Telangana, India
  • Google Scholar







 Received: 09 November 2016  Accepted: 08 December 2016  Published: 21 March 2017

Copyright © 2017 Author(s) retain the copyright of this article.
This article is published under the terms of the Creative Commons Attribution License 4.0


Proteases constitute most important enzymes owing to their wide variety of functions and have immense applications in various industries viz., medical, pharmaceutical, biotechnology, leather, detergent, and food industries. Despite of their wide spread occurrence in various sources, microorganisms present remarkable potential for proteolytic enzymes production due to their extensive biochemical diversity and susceptibility to genetic manipulation. The present study was aimed at isolating alkaline protease producing fungal members from dairy form effluents, designing the process variables for maximizing the protease production and determining the fibrinolytic potential of the partially purified alkaline protease. To achieve the specified objectives, the dairy form effluent was processed for the isolation of proteolytic fungi using suitable microbiological medium. All the fungal isolates were screened for their protease producing ability and the isolate showing highest alkaline protease production was selected for further studies. Optimization of different fermentative variables like carbon, nitrogen sources, pH, temperature and incubation period were carried out to enhance enzyme production. Ammonium sulphate fractionation was employed to partially purify the enzyme following which its fibrinolytic potential was determined. Based on morphological and microscopic studies, the selected fungal isolate was identified as Aspergillus niger. Optimization studies using OVAT (one variable at a time) method revealed an enhanced protease production in the presence of fructose as additional carbon source and ammonium sulphate as nitrogen source. The optimum incubation period, temperature and pH for enzyme production by the selected fungal isolate was found to be 92 h, 50°C and 10, respectively. The partially purified alkaline protease was efficient in the removal of blood stains emphasizing its fibrinolytic ability. An alkaline protease producing Fungal sp. was screened and isolated from dairy form effluent and it was found to be efficient in the removal of blood stains proving its fibrinolytic potential. Enzymes produced from microorganisms that can survive under extremes of pH could be particularly useful for commercial applications under high alkaline conditions.

 

Key words: Alkaline protease, dairy form effluent, optimization, Aspergillus niger, fibrinolytic potential.

Abidi F, Aissaoui N, Lazar S, Nejib M, Marzouki (2014). Purification and biochemical characterization of a novel alkaline protease from Aspergillus niger. Use in Antioxidant peptides production. J. Mater. Environ. Sci. 5:1490-1499.

 

Adinarayana K, Ellaiah P, Prasad DS (2003). Purification and partial characterization of thermostable serine alkaline protease from a newly isolated Bacillus subtilis PE-11. AAPS Pharm. Sci. Technol. 4:440-448.
Crossref

 
 

Al-Qodah Z, Daghistani H, Alananbeh K (2013). Isolation and characterization of thermostable protease producing Bacillus pumilus from thermal spring in Jordan. Afr. J. Microbiol. Res. 7(29):3711-3719.

 
 

Amrita Raj, Nancy K, Namrata P, Sourav B, Arijit D, Subbaramiah SR (2012). Enhancement of protease production by Pseudomonas aeruginosa isolated from dairy effluent sludge and determination of its fibrinolytic potential. Asian. Pac. J. Trop. Biomed. 2(3):S1845-S1851.
Crossref

 
 

Anshu G, Khare SK (2007). Enhanced production and characterization of a solvent stable protease from solvent tolerant Pseudomonas aeruginosa. Enzyme Microb. Technol. 42:11-16.
Crossref

 
 

Balaji N, Rajasekaran KM, Kanipandian N, Vignesh V, Thirumurugan R (2012). Isolation and screening of proteolytic bacteria from freshwater fish Cyprinus carpio. Int. Multidiscip. Res. J. 2:56-59.

 
 

Bhattacharya S, Bhardwaj S, Das A, Anand S (2011). Utilization of sugarcane bagasse for solid- state fermentation and characterization of α- amylase From Aspergillus flavus isolated from Muthupettai Mangrove, Tamil Nadu, India. Aust. J. Basic Appl. Sci. 5: 1012-1022.

 
 

Chandrasekaran S, Kumaresan SSP, Manavalan M (2015). Production and Optimization of Protease by Filamentous Fungus Isolated from Paddy Soil in Thiruvarur District Tamilnadu. J. Appl. Biol. Biotechnol. 3:66-69.
Crossref

 
 

Coral G, Arikan B, Ünaldi MN, Güvenmez H (2003). Thermostable alkaline protease produced by an Aspergillus niger strain. Ann. Microbiol. 53:491-498.

 
 

Deng A, WU J, Zhang Y, Zhang G, Wen T (2010). Purification and characterization of a surfactant-stable high-alkaline protease from Bacillus sp. B001. Bioresour. Technol. 101:7100-7116.
Crossref

 
 

Denizci AA, Kazan D, Abeln ECA, Erarslan A (2004). Newly isolated Bacillus clausii GMBAE 42: an alkaline protease producer capable to grow under highly alkaline conditions. J. Appl. Microbiol. 96:320-327.
Crossref

 
 

Hoog GS, Guarro J (2000). Explanatory chapters and keys to the genera, second ed. in: Hoog GS, Guarro J, Gene J, Figueras MJ (Eds.), Atlas of Clinical Fungi. Centraal bureau voor schimmel cultures. Spain Press., Netherlands and Universitat Rovira i Virgili. Pp.361-1008.

 
 

Ibrahim ASS, Al-Salamah AA (2009). Optimization of media and cultivation conditions for alkaline protease production by alkaliphilic Bacillus halodurans. Res. J. Microbiol. 4:251-259.
Crossref

 
 

Jellouli K, Bougatef A, Manni L, Agrebi R, Siala R, Younes I, Nasri M (2009). Molecular and biochemical characterization of an extracellular serine-protease from Vibrio etschnikovii J1. J. Ind. Microbiol. Biotechnol. 36:939-948.
Crossref

 
 

Johnvesly B, Naik GR (2001). Studies on the production of thermostable alkaline protease from thermophilic and alkaliphilic Bacillus sp. JB-99 in a chemical defined medium. Process Biochem. 37:139-144.
Crossref

 
 

Kalaiarasi K, Sunitha PU (2009). Optimization of alkaline protease production from Pseudomonas fluorescens isolated from meat waste contaminated soil. Afr. J. Biotechnol. 8:7035-7041.

 
 

Kalpana Devi M, Rasheedha Banu A, Gnanaprabhal GR, Pradeep BV, Palaniswamy M (2008). Purification, characterization of alkaline protease enzyme from native isolate Aspergillus niger and its compatibility with commercial detergents. Indian J. Sci. Technol. 1:1-6.

 
 

Kumar DJM, Venkatachalam P, Govindarajan N, Balakumaran MD, Kalaichelvan PT (2012). Production and purification of alkaline protease from Bacillus sp. MPTK 712 isolated from dairy sludge. Glob. Vet. 8:433-439.

 
 

Mala M, Srividya S (2010). Partial purification and properties of a laundry detergent compatible alkaline protease from a newly isolated Bacillus species Y. Indian. J. Microbiol. 50:309-317.
Crossref

 
 

Malathi S, Chakraborty R (1991). Production of Alkaline Protease by a New Aspergillus flavus Isolate under Solid-Substrate Fermentation Conditions for Use as a Depilation Agent. Appl. Environ. Microbiol. 57:712-716.

 
 

Mostafa El-Sayed E, Moataza MS, Hassan MA, Mohsen HS, Hassan HM (2012). Optimization conditions of extracellular proteases production from a newly isolated Streptomyces pseudogrisiolus NRC-15. E- J. Chem. 9:949-961.
Crossref

 
 

Mukhtar H, Ikram-Ul-Haq (2009). Production of Acid Protease by Aspergillus niger Using Solid State Fermentation. Pakistan J. Zool. 41:253-260.

 
 

Nehra KS, Dhillon S, Kamala Chowdhary, Randhin singh (2002). Production of alkaline protease by Aspergillus sp under submerged and solid state fermentation. Indian J. Microbial. 42:43-47.

 
 

Oyeleke SB, Egwim EC, Auta SH (2010). Screening of Aspergillus flavus and Aspergillus fumigatus strains for extracellular protease enzyme production. J. Microbiol. Antimicrob. 2:83-87.

 
 

PalaniveL P, Ashokkumar L, Balagurunathan R (2013). Production, purification and fibrinolytic characterization of alkaline protease from extremophilic soil fungi. Int. J. Pharm. Bio. Sci. 4:(B)101-110.

 
 

Radha S, Nithya VJ, Himakiran Babu R, Sridevi A, Prasad NBL, Narasimha B (2011). Production and optimization of acid protease by Aspergillus spp under submerged fermentation. Arch. Appl. Sci. Res. 3:155-163.

 
 

Rajkumar R, Jayappriyan KR, Kannan RP, Rengasamy R (2010). Optimization of culture conditions for the production of protease from Bacillus megaterium. J. Ecobiotechnol. 2:40-46.

 
 

Raju K, Jaya R, Ayyanna C (1994). Hydrolysis of casein by bajara protease importance. Biotechnol. Coming Decadea. 181:55-70.

 
 

Reese ET, Siu RGH, Levinson HS (1950). The biological degradation of cellulose derivatives and its regulation to the mechanism of cellular hydrolysis. J. Bacteriol. 59:485-489.

 
 

Rupali D (2015). Screening and Isolation of Protease Producing Bacteria from Soil Collected from Different Areas of Burhanpur Region (MP) India. Int.J.Curr.Microbiol. Appl. Sci. 4:597-606.

 
 

Sehar S, Hameed A (2011). Extracellular alkaline protease by a newly isolated halophilic Bacillus sp. Glob. J. Biotechnol. Biochem. 6:142-148.

 
 

Sharma AK, Sharma V, Jyoti S, Yadav B, Alam A, Prakash A (2015). Isolation and Screening of Extracellular Protease Enzyme from Bacterial and Fungal Isolates of Soil. Int. J. Sci. Res. Environ. Sci. 3:334-340.
Crossref

 
 

Singh SK, Singh SK, Tripathi VR, Khare SK, Garg SK (2011). Comparative one-factor-at-a-time, response surface (statistical) and benchscale bioreactor level optimization of thermoalkaline protease production from a psychrotrophic Pseudomonas putida SKG-1 isolate. Microb. Cell. Fact. 10:114-127.
Crossref

 
 

Sonia Sethi, Saksham Gupta (2015). Optimization of protease production from fungi isolated from soil. Int. J. Appl. Biol. Pharm. Tech. 6:149-154.

 
 

Tiwari ON, Bidyababy Devi T, Sarabati Devi K, Oinam G, Indrama T, Ojit K, Avijeet O, Ningshen L (2015). Isolation and optimization of alkaline protease producing Bacteria from undisturbed soil of NE-region of India falling under Indo-Burma biodiversity hotspots. J. Appl. Biol. Biotechnol. 3:25-31.
Crossref

 
 

Udandi Boominadhan, Rajendran Rajakumar, Palanivel Karpaga, Vinayaga Sivakumaar, Manoharan Melvin Joe (2009). Optimization of Protease Enzyme Production Using Bacillus Sp. Isolated from Different Wastes. Bot. Res. Int. 2:83-87.

 
 

Vadlamani S, Parcha SR (2011). Studies on industrially important alkaline protease production from locally isolated superior microbial strain from soil microorganisms. Int. J. Biotechnol. Appl. 3:102-105.

 
 

Vaishali Choudhary, Jain PC (2012). Optimization of process parameters for alkaline protease production by Aspergillus versicolor PF/F/107. J. Acad. Indus. Res. 1:8-14.

 
 

Vishwanatha T, Jain SN, Reena V, Divyashree BC, Siddalingeshwara KG, Karthic J, Sudipta KM (2010). Screening of substrates for protease production from Bacillus licheniformis. Int. J. Eng. Sci. Technol. 2:6550-6554.

 

 


APA Lanka, S., Anjali, CH. & Pydipalli, M. (2017). Enhanced production of alkaline protease by Aspergillus niger DEF 1 isolated from dairy form effluent and determination of its fibrinolytic ability. African Journal of Microbiology Research, 11(11), 440-449.
Chicago Suseela Lanka, CH. Anjali and Muralidhar Pydipalli. "Enhanced production of alkaline protease by Aspergillus niger DEF 1 isolated from dairy form effluent and determination of its fibrinolytic ability." African Journal of Microbiology Research 11, no. 11 (2017): 440-449.
MLA Suseela Lanka, CH. Anjali and Muralidhar Pydipalli. "Enhanced production of alkaline protease by Aspergillus niger DEF 1 isolated from dairy form effluent and determination of its fibrinolytic ability." African Journal of Microbiology Research 11.11 (2017): 440-449.
   
DOI 10.5897/AJMR2016-8379
URL http://www.academicjournals.org/journal/AJMR/article-abstract/512797E63282

Subscription Form