RESEARCH PAPER
Enzyme-mediated protoplast formation of Cuninghamella echinulata – preliminary studies
1
Wrocław University of Technology, Faculty of Chemistry, Wrocław, Poland
Submission date: 2015-12-10
Final revision date: 2016-02-15
Acceptance date: 2016-02-15
Publication date: 2016-04-20
BioTechnologia 2016;97(1):27-32
KEYWORDS
TOPICS
ABSTRACT
In this report, we have presented the analysis of the enzymatic methods of protoplasts formation of Cunninghamella echinulata. Hydrolytic enzymes of different origin (i.e. prokaryotic and eukaryotic) such as chitinase (EC.3.2.1.14), lyticase (mix of lytic enzymes), and proteinase K (EC 3.4.21.64) were used for fungal cell wall disruption. An effective protoplast formation was achieved with the following enzyme solutions: chitinase (0.33 U A ml!1), proteinase K (1.8-3.6 U A ml!1), and lyticase (160 UA ml!1). Fluorescent microscopy was used to evaluate the ability of the tested enzymes to effectively produce protoplasts.
REFERENCES (31)
1.
Ahmed M., Barkly E.I. (2006) Gene transfer between different Trichoderma species and Aspergillus niger through intergeneric protoplast fusion to convert ground rice straw to citric acid and cellulose. Appl. Biochem. Biotech. 135: 117-132.
2.
Alvarez E., Sutton D.A., Cano J., Fothergill A.W., Stchigel A., Rinaldi M.G., Guarro J. (2009) Spectrum of Zygomycete species identified in clinically significant specimens in the United States. J. Clin. Microbiol. 47: 1650-1656.
3.
Andrade V.S., Neto B.B., Souza W., Campos-Takaki G.M. (2000) A factorial design analysis of chitin production by Cunninghamella elegans. Can. J. Microbiol. 46: 1042-1045.
4.
Asha S., Vidyavathi M. (2009) Cunninghamella – a microbial model for drug metabolism studies – a review. Biotechnol. Adv. 27: 16-29.
5.
Avram D., Petcu I., Radu M., Dan F., Stan R. (1992) Electrically induced protoplast fusion for ergosterol producing yeast strain improvement. J. Basic. Microbiol. 32: 369-372.
6.
Cerniglia C.E. (1992) Biodegradation of polycyclic aromatic hydrocarbons. Biodegradation 3: 351-368.
7.
Diaz-Jimenez D.F., Perez-Garcia L.A., Martinez-Alvarez J.A., Mora-Montes H.M. (2012) Role of the fungal cell wall in pathogenesis and antifungal resistance. Curr. Fungal Infect. Rep. 6: 275-282.
8.
El-Morsy E.S.M. (2004) Cunninghamella echinulata a new biosorbent of metal ions from polluted water in Egypt. Mycologia 96: 1183-1189.
9.
Fakas S., Galiotou P.M., Papanikolaou S., Komaitis M., Aggelis G. (2007) Compositional shifts in lipid fractions during lipid turnover in Cuninghamella echinulate. Enzym. Microbiol. Technol. 40: 1321-1327.
10.
Feofilova E.P. (2010) The fungal cell wall: Modern concepts of its composition and biological function. Microbiology 79: 711-720.
11.
Gema H., Kavadia A., Dimou D., Tsagou V., Komaitis M., Aggelis G. (2002) Production of γ-linolenic acid by Cuninghamella echinulata cultivated on glucose and orange peel. Appl. Microbiol. Biotechnol. 58: 303-307.
12.
Hartl L., Zach S., Seidl-Seiboth V. (2012) Fungal chitinases: diversity, mechanistic properties and biotechnological potential. Appl. Microbiol. Biotechnol. 93: 533-543.
13.
Holland H.L. (1999) Recent advances in applied and mechanistic aspects of the enzymatic hydroxylation of steroids by whole-cell biocatalyst. Steroids 64: 178-186.
14.
Kavadia A., Komaitis M., Chevalot I., Blanchard F., Marc I., Aggelis G. (2001) Lipid and γ-linolenic acid accumulation in strains of zygomycetes growing on glucose. J. Am. Oil Chem. Soc. 78: 341-346.
15.
Kirimura K., Lee S.P., Nakajima I., Kawabe S.M., Usami S. (1988) Improvement in citric acid production by haploidization of A. niger diploid strains. J. Ferment. Technol. 66: 375-382.
16.
Koushki M.M., Rouhani R., Farsi M. (2011) Genetic manipulation of fungal strains for the improvement of heterologous genes expression (a mini-review). Afr. J. Biotechnol. 10: 7939-7948.
17.
Lunin V.V., Sergeeva Y.E., Galanina L.A., Mysyakina I.S., Ivashechkin A.A., Bogdan V.I., Feofilova E.P. (2013) Biodiesel fuel production from lipids of filamentous fungi. Appl. Biochem. Microb. 49: 46-52.
18.
Nwe N., Stevens W.F., Tokura S., Tamura H. (2008) Characterization of chitosan and chitosan-glucan complex extracted from the cell wall of fungus Gongronella butleri USDB 0201 by enzymatic method. Enzym. Microbiol. Technol. 42: 242-250.
19.
Papanikolau S., Panayotou M.G., Fakas S., Komaitis M., Aggelis G. (2007) Lipid production by oleaginous mucorales cultivated on renewable carbon source. Eur. J. Lipid Sci. Tech. 109: 1060-1070.
20.
Pasha C., Kuhad R.C., Rao L.V. (2007) Strain improvement of termotolerant Saccharomyces cerevisiae VS3 strain for better utilization of lignocellulosic substrates. J. App. Microbiol. 103: 1480-1489.
21.
Pękala E., Kubowicz P., Łażewska D. (2012) Cunninghamella as a microbiological model for metabolism of histamine H3 receptor antagonist 1-[3-(4-tert-butylphenoxy)propyl] piperidine. Appl. Biochem. Biotech. 168: 1584-1593.
22.
Scott C.T., Vergoignan C., Feron G., Durand A. (2001) Glucosamine measurement as indirect method for biomass estimation of Cunninghamella elegans grown in solid state cultivation conditions. Biochem. Eng. J. 7: 1-5.
23.
Seigle-Murandi F.M., Krivobok S.M.A., Steiman R.L., Benoit- Guyod J.L.A., Thiault G.A. (1991) Biphenyl Oxide Hydroxylation by Cunninghamella echinulate. J. Agric. Food Chem. 39: 428-430.
24.
Sorbotten A., Horn S.J., Eijsink V.G.H., Varum K.M. (2005) Degradation of chitosans with chitinase B from Serratia marcescens. Production of chito-oligosacharides and insight into enzyme processivity. FEBS J. 272: 538-549.
25.
Sotnikova I.V., Telesnina C.N., Lebed E.S., Dmitrieva S.V., Zhukov V.G. (1988) Antibiotic production in variants of Penicillium chrysogenum isolated after protoplasting stage and exposure to mutants at the protoplasting stage. Antibiot. Chemot. 33: 211-217.
26.
Srisilam K., Veeresham C. (2003) Biotransformation of drugs by microbial cultures for predicting mammalian drug metabolism. Biotechnol. Adv. 21: 3-39.
27.
Stamford T.C.M., Stamford T.L.M., Stamford N.P., Neto B.B., Campos-Takaki G.M. (2007) Growth of Cunninghamella elegans UCP 542 and production of chitin and chitosan using yam bean medium. Electron. J. Biotechnol. 10: 61-68.
28.
Tan S.C., Tan T.K., Wong S.M., Khor E. (1996) The chitosan yield of Zygomycetes at their optimum harvesting time. Carbohyd. Polym. 30: 239-242.
29.
Verma N., Bansal M.C., Kumar V. (2008) Protoplast Fusion technology and its biotechnological applications. Chem. Eng. Trans. 14: 113-120.
30.
Zhang D., Yang Y., Leakey J.E., Cerniglia C.E. (1996) Phase I and phase II enzymes produced by Cunninghamella elegans for the metabolism of xenobiotics. FEMS Microbiol. Lett. 138: 221-226.
| eISSN: | 2353-9461 |
| ISSN: | 0860-7796 |
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