Research Article | Volume: 4, Issue: 4, July-August, 2016

Production and Characterization of Collagenase by Penicillium sp. UCP 1286 Isolated From Caatinga Soil

Maria Carolina de Albuquerque Wanderley Jose Manoel Wanderley Duarte Neto Carolina de Albuquerque Lima Sara Isabel da Cruz Silverio Jose Luiz de Lima Filho Jose Antonio Couto Teixeira Ana Lucia Figueiredo Porto   

Open Access   

Published:  Aug 26, 2016

DOI: 10.7324/JABB.2016.40401

A new Penicillium sp. strain isolated from the soil of Caatinga, a Brazilian Biome (UCP 1286) was selected for collagenase production. Fermentation system allowing obtention of collagenolytic activity about 2.7 times higher than existing data, with the highest values of collagenolytic and specific activity (379.80 U/mL, 1460.77 U/mg, respectively), after 126 hours. Applying a factorial design, enzyme production was increased by about 65% compared to the preliminary results. The factorial design demonstrated the existence of two factors with statistical significance on the production of the enzyme: pH and temperature, both with negative effects. Enzyme was found to be more active at pH 9.0 and 37 °C, and also to be very stable in comparison with the collagenase produced by other microorganisms. The enzyme seems to belong to collagenolytic serine proteases family. Concerning the substrate specificity, it was observed that the highest enzyme activity corresponds to azocoll, there was no relevant activity on azocasein and the enzyme showed to be more specific to type V collagen and gelatin than the commercial colagenase produced by Clostridium histolyticum. Major band observed at electrophoresis was approximately 37 kDa. Zymogram analysis confirmed the collagenolytic activity. All data indicates this enzyme as promising biotechnology product.

Keyword:     Collagenolytic Enzymes Factorial Design Fermentation Filamentous Fungi Specificity.


Wanderley MCA, Neto JMWD, Lima CA, Silverio SIC, Filho JLL, Teixeira JAC, Porto ALF. Production and Characterization of Collagenase by Penicillium sp. UCP 1286 Isolated From Caatinga Soil. J App Biol Biotech. 2016; 4 (04): 001-010. DOI: 10.7324/JABB.2016.40401

Copyright: Author(s). This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike license.

HTML Full Text

1. Jain R, Jain PC. Production and partial characterization of collagenase of Streptomyces exfoliatus CFS 1068 using poultry feather. Indian J Exp Biol 2010;48:174-8.

2. Suphatharaprateep W, Cheirsilp B, Jongjareonrak A. Production and properties of two collagenases from bacteria and their application for collagen extraction. N Biotechnol 2011;28:649-55.

3. Di Lullo G a., Sweeney SM, Körkkö J, Ala-Kokko L, San Antonio JD. Mapping the ligand-binding sites and disease-associated mutations on the most abundant protein in the human, type I collagen. J Biol Chem 2002;277:4223-31.

4. Müller WEG. The origin of metazoan complexity: porifera as integrated animals. Integr Comp Biol 2003;43:3-10.

5. Tran LH, Nagano H. Isolation and Characteristics of Bacillus subtilis CN2 and its Collagenase Production. Food Microbiol Saf 2002;67:3-6.

6. Wu Q, Li C, Li C, Chen H, Shuliang L. Purification and characterization of a novel collagenase from Bacillus pumilus Col-J. Appl Biochem Biotechnol 2010;160:129-39.

7. Dresden MH. Evidence for the role of collagenase in collagen metabolism. Nature 1971.

8. Lima CA, Viana Marques DA, Neto BB, Lima Filho JL, Carneiro-da-Cunha MG, Porto ALF. Fermentation medium for collagenase production by Penicillium aurantiogriseum URM4622. Biotechnol Prog 2011;27:1470-7.

9. Lima CA, Filho JLL, Neto BB, Converti A, Carneiro da Cunha MG, Porto ALF. Production and characterization of a collagenolytic serine proteinase by Penicillium aurantiogriseum URM 4622: A factorial study. Biotechnol Bioprocess Eng 2011;16:549-60.

10. Lima CA, Júnior ACVF, Filho JLL, Converti A, Marques D a. V, Carneiro-da-Cunha MG, et al. Two-phase partitioning and partial characterization of a collagenase from Penicillium aurantiogriseum URM4622: Application to collagen hydrolysis. Biochem Eng J 2013;75:64-71.

11. Ravanti L, Kahari VM. Matrix metalloproteases in wound repair. Int J Mol Med 2000;6:391-407.

12. Shanmughapriya S, Kiran GS, Natarajaseenivasan K. Optimization of extracellular thermotolerant alkaline protease produced by marine Roseobacter sp. (MMD040). Bioproc Biosys Eng 2008;31:427-33.

13. Barret AJ, Rawlings ND, Woessner JF. Handbook of Proteolytic Enzymes. 2nd. ed. Oxford, U. K.: 2004.

14. Hamdy HS. Extracellular collagenase from Rhizoctonia solani : Production , purification and characterization. Indian J Biotechnol 2008;7:333-40.

15. Rao MB, Tanksale a M, Ghatge MS, Deshpande V V. Molecular and biotechnological aspects of microbial proteases. Microbiol Mol Biol Rev 1998;62:597-635.

16. Daboor SM, Budge SM, Ghaly AE, Brooks S, Dave D. Extraction and Purification of Collagenase Enzymes : A Critical Review. Am J Biochem Biotechnol 2010;6:239-63.

17. Mookthiar K, Steinbrink SD, Van Wart HE. Mode of hydrolysis of collagen-like peptidase by class I and class II Clostridium hystolyticum collagenases: Evidence for both endopeptidase and tripeptidyl-carboxypeptidase activities. Biochemistry1 985;24:6527-33.

18. Peterkofsky B. Bacterial collagenase. Methods Enzym 1982;82:453-71.

19. Sandhya C, Sumantha A, Szakacs G, Pandey A. Comparative evaluation of neural protease production by Aspergillus oryzae in submerged and solid state fermentation. Process Biochem 2005;40:2689-94.

20. Yakovleva MB, Khoang TL, Nikitina ZK. Collagenolytic activity in several species of deuteromycetes under various storage conditions. Appl Biochem Microbiol 2006;42:431-4.

21. de Siqueira ACR, da Rosa NG, Motta CMS, Cabral H. Peptidase with keratinolytic activity secreted by Aspergillus terreus during solid-state fermentation. Brazilian Arch Biol Technol 2014;57:514-22.

22. Rosso BU, Lima CDA, Porto TS, de Oliveira Nascimento C, Pessoa A, Converti A, et al. Partitioning and extraction of collagenase from Penicillium aurantiogriseum in poly(ethylene glycol)/phosphate aqueous two-phase system. Fluid Phase Equilib 2012;335:20-5.

23. Lima CA, Rodrigues PMB, Porto TS, Viana D a., Lima Filho JL, Porto ALF, et al. Production of a collagenase from Candida albicans URM3622. Biochem Eng J 2009;43:315-20.

24. Viani FC, Cazares Viani PR, Gutierrez Rivera IN, da Silva ÉG, Paula CR, Gambale W. Actividad proteolítica extracelular y análisis molecular de cepas de Microsporum canis aisladas de gatos con y sin sintomatología. Rev Iberoam Micol 2007;24:19-23.

25. Ikram-Ul-Haq H, Mukhtar. Biosynthesis of acid proteases by Penicillium griseoroseum IH-02 in solid-state fermentation. Pakist J Bot 2007;39:2717-24.

26. Menezes R, Sampaio E, Giongo V, Pérez-Marin A. Biogeochemical cycling in terrestrial ecosystems of the Caatinga Biome. Brazilian J Biol 2012;72:643-53.

27. Ferreira ACC, Leite LFC, de Araújo ASF, Eisenhauer N. Land-Use Type Effects on Soil Organic Carbon and Microbial Properties in a Semi-Arid Region of Northeast Brazil. L Degrad Dev 2014;178:n/a - n/a.

28. Pacchioni RG, Carvalho FM, Thompson CE, Faustino ALF, Nicolini F, Pereira TS, et al. Taxonomic and functional profiles of soil samples from Atlantic forest and Caatinga biomes in northeastern Brazil. Microbiologyopen 2014;3:299-315.

29. Chavira RJ, Burnett TJ, Hageman JH. Assaying Proteinases with Azocoll. Anal Biochem 1984;136:446-50.

30. Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 1976;72:248-54.

31. Endo A, Murakawa S, Shimizu H, Shiraishi Y. Purification and properties of collagenase from Streptomyces species. J Biochem 1987;102:163-70.

32. Rosen H. A modified ninhydrin colorimetric analysis for amino acids. Arch Biochem Biophys 1975;67:10-5.

33. Moore S, Stein WH. Photometric ninhydrin method for use in the chromatography of amino acids. J Biol Chem 1948;176:367-88.

34. Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970;227:680-5.

35. Baehaki A, Sukamo, Syah D, Setyahadi S, Suhartono MT. Production and characterization of collagenolytic protease from Bacillus licheniformis F11.4 originated from Indonesia. Asian J Chem 2014;26:2861-4.

36. Mahmoud Y a.-G, Abu El-Souod SM, El-Shourbagy SM, El-Badry ASM. Characterisation and inhibition effect of cetrimide on collagenase produced by Aspergillus flavus, isolated from mycotic ulcers. Ann Microbiol 2007;57:109-13.

37. Ok T, Hashinaga F. Detection and production of extracellular collagenolytic enzyme from Zygosaccharomyces rouxii. J Gen Appl Microbiol 1996;42:517-23.

38. Chellapan S, Jasmin C, Basheer SM, Elyas K, Bhat SG, Chandrasekaran M. Production, purification and partial characterization of a novel protease from marine Engyodontium album BTMFS10 under solid station fermentation. Process Biochem 2006;41:956-61.

39. Sakurai Y, Inoue H, Nishii W, Takahashi T, Iino Y, Yamamoto M, et al. Purification and characterization of a major collagenase from Streptomyces parvulus. Biosci Biotechnol Biochem 2009;73:21-8.

40. Adachi E, Hayashi T. “In vitro” formation of fine fibrils with a D-periodic banding pattern type V collagen. Relat Res 1985;5:225-32.

41. Kiriviko KI, Mlyllyla R. Biosynthesis of the collagens. In: Piez KA, Reddi AH, organizadores. Extracell. Matrix Biochem., New York: Elsevier; 1984, p. 83-118.

42. Moreland LW, Gay RE, Gay S. Collagen autoantibodies in patients with vasculitis and systemic lupus erythematosus. Clin Immunol Immunopathol 1991;60:412-218.

43. James GT. Inactivation of the protease inhibitor phenylmethylsulfonyl fluoride in buffers. Anal Biochem 1978;86:574-9.

44. Baehaki A, Suhartono MT, Syah D, Sitanggang AB, Setyahadi S, Meinhardt F. Purification and characterization of collagenase from Bacillus licheniformis F11.4. African J Microbiol Res 2012;6:2373-9.

45. Roy P, Colas B, Durand P. Purification, kinetical and molecular characterizations of a serine collagenolytic protease from greenshore crab (Carcinus maenas) digestive gland. Comp Biochem Physiol 1994;36:247-301.

46. Matsushita K, Toyama H, Adachi O. Respiratory chains and bioenergetics of acetic acid bacteria. Adv Microb Physiol 1994;36:247-301.

47. Kristjánsson MM, Gudmundsdóttir S, Fox JW, Bjamason JB. Characterization of collagenolytic serine proreinase from the Atlantic cod (Gadus morhua). Comp Biochem Physiol 1995;110:707-17.

48. Sivakumar P, Sampath P, Chandrakasan G. Collagenolytic metalloprotease (gelatinase) from the hepatopancreas of the marine crab, Scylla serrata. Comp Biochem Physiol 1999;123:273-9.

Article Metrics
42 Views 119 Downloads 161 Total



Related Search

By author names

Similar Articles

Enzymes and qualitative phytochemical screening of endophytic fungi isolated from Lantana camara Linn. Leaves

Mbouobda Hermann Desire , Fotso Bernard , Muyang Rosaline Forsah , Chiatoh Thaddeus Assang, Omokolo Ndoumou Denis

Impact of Phyllanthus amarus extract on antioxidant enzymes in Drosophila melanogaster

N. Manasa, J. S. Ashadevi

Physiological and biochemical characterization of Sesamum germplasms tolerant to NaCl

Tapaswini Hota, C. Pradhan, G. R. Rout

Enzymatic responses of Clarias gariepinus (Burchell, 1822) exposed to sub-lethal concentrations of an oilfield wastewater

Nedie Patience Akani, Ugwemorubon Ujagwung Gabriel

Effect of various dietary fats supplementation on the liver glycogen, protein and digestive enzymes activities in striped murrel, Channa striatus

Rajesh Dayal, Prem Prakash Srivastava , Joykrushna Jena, Sudhir Raizada, Akhilesh Kumar Yadav, Anita Bhatnagar, Shipra Chowdhary

Trichoderma oligosaccharides priming mediates resistance responses in pearl millet against downy mildew pathogen

Boregowda Nandini, Puttaswamy Hariprasad, Harischandra Sripathy Prakash, Nagaraja Geetha

Morphological, enzymatic screening, and phylogenetic analysis of thermophilic bacilli isolated from five hot springs of Myagdi, Nepal

Punam Yadav , Suresh Korpole, Gandham S Prasad, Girish Sahni , Jyoti Maharjan, Lakshmaiah Sreerama, Tribikram Bhattarai

Cold-active enzymes in food biotechnology: An updated mini review

Mohammed Kuddus

Aroclor 1254 induced oxidative stress and histopathological changes in mice liver

Jalpa Raja, Shweta Pathak, Rahul Kundu

The effects of diet and temperature on enzymes, hormones, and fecundity of the African Catfish Clarias gariepinus (Burchell 1822)

Waleed Abdul-Aziz A. Al-Deghayem, El Amin Mohamed Suliman

A study on the salinity stress effects on the biochemical traits of seedlings and its relationship with resistance toward sensitive and tolerant flax genotypes

Yousef Alaei, Seyed Kamal Kazemitabar, Mohammad Zaefi Zadeh, Hamid Najafi Zarini, Gaffar Kiani

Role of glutathione reductase and catalase enzyme in antioxidant defense mechanism in controlling fluoride-induced oxidative stress

Komal Sharma, Mamta Choudhary, Khushbu Verma

Dye degradation potential and its degradative enzymes synthesis of Bacillus cereus SKB12 isolated from a textile industrial effluent

Thangaraj Sheela, Senthil Kumar Sadasivam

Qualitative and quantitative analysis of Precocene II, estimation of enzymatic, nonenzymatic antioxidant, and cytotoxic potentials of methyl jasmonate-elicited shoot culture of Ageratum conyzoides Linn.

Selvaraj Vasantharani, Ramaraj Thirugnanasampandan, Gunasekaran Bhuvaneswari

Isolation and characterization of starch degrading bacteria from disparate soil samples

Vidhyutha Srivathsan, Mahima Bhandari, Priya Swaminathan

In silico modeling, docking of ThPON1-like protein, and in vitro validation of pesticide tolerance in Trichoderma harzianum

Archana Kumari, Krishna Sundari Sattiraju

Endophytic Fungi as Emerging Bioresources for Bioactive Compounds for Sustainable Development

Divjot Kour, Neelam Yadav, Ajar Nath Yadav

Microbes mediated plastic degradation: A sustainable approach for environmental sustainability

Harpreet Kour, Sofia Shareif Khan, Divjot Kour, Shafaq Rasool, Yash Pal Sharma, Pankaj Kumar Rai, Sangram Singh, Kundan Kumar Chaubey, Ashutosh Kumar Rai, Ajar Nath Yadav

Assessment of morpho-agronomic and yield attributes in gamma-irradiated mutants of Kalanamak rice (Oryza sativa L.)

Tanmai Mishra, Anjali Singh, Virendra Kumar Madhukar, Ashutosh Kumar Verma, Shambhavi Mishra, Rajveer Singh Chauhan

Phenotypic and genotypic detection of Beta-lactamase producing Pseudomonas aeruginosa isolated from Haryana, India

Khem Chand,, Amit Lath, Aparna Yadav, Madhu Sharma, Nater Pal Singh, Sanjay Kumar, Rajiv Yadav, Sawraj Singh, Anita Rani Santal

Application of two-level factorial design in optimization of maceration extraction of Curcuma longa curcuminoids extracts

Rudi Heryanto,, Siti Sa’diah, Waras Nurcholis,

Effect of Wild and Mutant Strain of Lasiodiplodia Pseudotheobromae Mass Produced on Rice Bran as a Potential Bioherbicide Agents for Weeds under Solid state Fermentation

Adetunji C. O.a, Oloke J.Kb

Medium Formulation and its optimization for increased protease production by Penicillium sp. LCJ228 and its potential in blood stain removal

V. Benluvankar, S. Evelyne Priya, J. Joel Gnanadoss

Screening, Selection and Optimization of the Culture Conditions for Tannase Production by Endophytic Fungi Isolated from Caatinga

Rayza Morganna Farias Cavalcanti, Pedro Henrique de Oliveira Ornela, João Atílio Jorge, Luís Henrique Souza Guimarães

Proteome and fermentative parameters of Saccharomyces cerevisiae CAT-1 under Very High Gravity Fermentation (VHGF) using sugarcane juice

Gabriela de Sá Azarias, Heloisy Suzes Barbosa, Cynthia Barbosa Rustiguel, José Cesar Rosa, José Roberto Ernandes, Luis Henrique Souza Guimarães

Production, characterization and optimization of fermented tomato and carrot juices by using Lysinibacillus sphaericus isolate

Naga Sivudu Seelam, Harika Akula, Umamahesh Katike, Vijaya Sarathi Reddy Obulam

Fermentative Production of Polyhydroxyalkanoates (PHAs) from Glycerol by Zobellella taiwanensis Azu-IN1

Mohamed Ali Abdel-Rahman, Said El-Sayed Desouky, Mohamed Salah Azab, Mahmoud E. Esmael

Biochemical and molecular characterization of a new pullulan producer Rhodosporidium paludigenum PUPY-06

RS Singh, Navpreet Kaur

Biocatalysis of agro-processing waste by marine Streptomyces fungicidicus strain RPBS-A4 for cellulase production

Rajanikanth Akurathi, Damodharam Thoti

Inulinase production in shake-flask fermentations from Mucor circinelloides BGPUP-9

Ram Sarup Singh, Kanika Chauhan, Randeep Kaur, Ramanpreet Kaur

Laccase production by Myrothecium gramineum and its optimization under solid state fermentation using cowpea pod as substrate

V. Daphne Vivienne Gnanasalomi, J. Joel Gnanadoss

Screening and identification of amylase producing strains of Bacillus

Kumar Pranay, Shree Ram Padmadeo, Vijay Jha, Birendra Prasad

Analysis of physicochemical and sensory parameters of wine produced from Carica papaya

Suhail Cholassery, Vidhu Krishna, SreethuVeliparambil Sethuraj,Shabnam Shajahan Rehina, Vandana Ranganathan, Lisha Chanassery Dileep, Kuzhunellil Raghavanpillai Sabu, Bhama Ramachandran Rajesh, Ramachandran Pratap Chandran

Enhanced biobutanol production by optimization of the medium from Clostridium acetobutylicum MTCC 11274 using macroalgae Gracilaria edulis

Shelly Rana, Kashyap Dubey, Ashwani Dhingra

Media optimization studies and production of adenosylcobalamin (Vitamin B12) by environment friendly organism Rhizobium spp

Neha Nohwar, Rahul V. Khandare, Neetin S. Desai

Amylase production by Aspergillus niger in submerged cultivation using cassava

Muralikandhan Kamaraj, Dhanasekaran Subramaniam

Production and purification of extracellular fungal cellulases using agricultural waste

Abishna Burugu, Dheerendra Kumar Suman, Chandrasekhar Chanda

Enhancement of Vitamin D2 content through ultraviolet-B irradiation in submerged cultivated Pleurotus eryngii mycelia using response surface methodology

Umesh Singh, Ashwani Gautam, Satyawati Sharma

Role of lacto-fermentation in reduction of antinutrients in plant-based foods

Mehak Manzoor, Deepti Singh, Gajender Kumar Aseri, Jagdip Singh Sohal, Shilpa Vij, Deepansh Sharma

Impact of diverse processing treatments on nutritional and anti-nutritional characteristics of soybean (Glycine max L.)

Priyanka Thakur, Krishan Kumar, Naseer Ahmed, Ajar Nath Yadav, Sunil Kumar, Qurat Ul Eain Hyder Rizvi, Divya Chauhan, Sumaira Jan

Effect of diverse fermentation treatments on nutritional composition, bioactive components, and anti-nutritional factors of finger millet (Eleusine coracana L.)

Sumaira Jan, Krishan Kumar, Ajar Nath Yadav, Naseer Ahmed, Priyanka Thakur, Divya Chauhan, Qurat-Ul-Eain Hyder Rizvi, Harcharan Singh Dhaliwal

Solid-state fermentation of groundnut (Arachis hypogaea) shell using Trichoderma sp., tape yeast, and tempeh yeast to produce cellulase

Muhammad Yusuf Abduh, Chalil Rizqullah Ramadhan, Alfanny Putri Fadhlilah, Siti Dhiffah Nabilah Abdul, Khairul Hadi Burhan

Identification and characterization of acidosis on in vitro rumen fermentation with feeds based on grass, rice bran, concentrate, and tofu pulp

Darwin, Tiya Humaira, Ami Muliawati

Impact of soaking, germination, fermentation, and roasting treatments on nutritional, anti-nutritional, and bioactive composition of black soybean (Glycine max L.)

Divya Chauhan, Krishan Kumar, Naseer Ahmed, Priyanka Thakur, Qurat Ul Eain Hyder Rizvi, Sumaira Jan, Ajar Nath Yadav

Extraction of a novel bacteriocin from Lacticaseibacillus casei VITCM05 and its antibacterial activity against major food-borne pathogens

Jannatul Firdous Siddique, Mohanasrinivasan Vaithilingam

Isolation, characterization and optimization of keratinolytic bacteria from chicken feather waste dumping site

Thiyagarajan Amuthavalli, Cyril Ravi

High cell density fermentation strategies for the production of bovine lactoferrin in Pichia pastoris

Thuy Thi Thu Trinh, Huong Thu Ngo, Phong Quoc Truong

Increasing polyphenol antioxidant in Orthosiphon stamineus Benth leaves with fermentation extraction by Saccharomyces cerevisiae ATCC-9763

Muhammad Aria Chandra, Khaswar Syamsu, Laksmi Ambarsari, Nurul Fatimah, Waras Nurcholis,

A novel fibrinolytic enzyme producer from mangrove soil sediments: Screening, isolation, strain improvement, and fermentation

Bhavana Sompalli, Alok Malaviya,,

Effects of process parameters on the alcoholic fermentation of pomelo (Citrus grandis (L.) Osbeck) juice

Huynh Xuan Phong, Tran Thi Yen Nhi, Nguyen Ngoc Thanh, Le Dang Truong

Recent advances in the processing of Napier grass (Pennisetum purpureum Schumach) as a potential bioenergy crop for bioethanol production

Priya Chamoli, Samiksha Jhildiyal, Palak Agrawal, Navin Kumar, Pallavi Singh

Optimizing solid-state fermentation for metabolite enrichment by Aspergillus tamarii on rice bran and wheat

Syaefudin Suminto,, Arthur Alba Huang, Uswatun Hasanah, Waras Nurcholis,

Statistical optimization of process variables for anti-hypercholesterolemic metabolites production from Monascus purpureus MTCC 369 fermented finger millet

Monu Kumar, Gaurav Chaudhary, Anita Rani Sehrawat, Anoop Singh, Guddu Kumar Gupta

Investigation of optimal parameters for aspartic protease production from Aspergillus terreus using solid-state fermentation

Sadeq Fenjan Hasnawi,, Alaa Jabbar Abd Al-Manhel, Zena Kadhim AL-Younis

The fermentation conditions of low alcoholic three-leaved (Cayratia trifolia (L.) Domin) cider using Saccharomyces cerevisiae HG1.3

Tien Thi Kieu Doan, Mi Thi Ngoc Huynh, Thu Thi Minh Tran, Thanh Huu Nguyen, Son Thi Bich Le, Thanh Ngoc Nguyen, Phong Xuan Huynh