Abstract
The purpose of this investigation was to study the effect ofStreptomyces marinensis NUV-5 cells immobilized in calcium alginate for the production of neomycin. The effect of various parameters, such as the effect of alginate concentration (1%, 2%, 3%, 4%, and 5% wt/vol), the effect of cation (caCl2, BaCl2, and SrCl2), the concentration of cation (0.01M, 0.125M, 0.25M, 0.375M, and 0.5M), the curing times (1, 6, 11, 16, and 21 hours), and the diameter of the bead (1.48, 2.16, 3.24, 4.46, and 5.44 mm), on neomycin production and bead stability were studied. The effect of maltose (4%, 3%, 2%, and 1% wt/vol) and sodium glutamate (0.6%, 0.3%, 0.15%, and 0.075%) wt/vol) concentration on neomycin production was also studied. Better neomycin production was achieved with optimized parameters, such as alginate at 2% wt/vol, 0.25M CaCl2, 1-hour curing time, and 3.24 mm bead diameter. Effective neomycin production was achieved with 3% wt/vol maltose and 0.6% wt/vol sodium glutamate concentration. The repeated batch fermentations were conducted (every 96 hours) using the optimized alginate beads, employing the production medium with 3% wt/vol maltose and 0.6% wt/vol sodium glutamate along with minerals salts solution. The increase in antibiotic production was observed up to the 5th cycle, and later gradual decrease in antibiotic production was observed. Comparison of the total antibiotic production with free cells and immobilized cells was also done. An enhanced antibiotic productivity of 32% was achieved with immobilized cells over the conventional free-cell fermentation, while 108% more productivity was achieved over the washed free-cell fermentation. From these results it is concluded that the immobilized cells ofS marinensis NUV-5 in calcium alginate are more efficient for the production of neomycin with repeated batch fermentation.
Keywords: neomycin production, Streptomyces marinensis NUV-5, immobilized cells
Full Text
The Full Text of this article is available as a PDF (259.9 KB).
References
- 1.Waksman SA, Lechvalier HA. Neomycin, a new antibiotic active against strep tomycin resistant bacteria, including tuberculosis organisms. Science. 1949;19:305–305. doi: 10.1126/science.109.2830.305. [DOI] [PubMed] [Google Scholar]
- 2.Sambamurthy K, Ellaiah P. A new Streptomycete producing neomycin (B&C) complex—S. marinensis (part I) Hindustan Antibiot Bullet. 1974;17:24–27. [PubMed] [Google Scholar]
- 3.Lechevalier HA. The 25 years of neomycin. CRC Crit Rev Microbiol. 1975;3(4):359–397. doi: 10.3109/10408417509108756. [DOI] [PubMed] [Google Scholar]
- 4.Dulmage HT. The production of neomycin byStreptomyces fradiae in synthetic media. Appl Microbiol. 1953;1:103–106. doi: 10.1128/am.1.2.103-106.1953. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 5.Sebek OK. Synthesis of neomycin C14 byStreptomyces fradiae. Arch Biochem Biophys. 1995;57:71–79. doi: 10.1016/0003-9861(55)90178-8. [DOI] [PubMed] [Google Scholar]
- 6.Majumdar MK, Majumdar SK. Utilization of carbon and nitrogen containing compounds for neomycin production byStreptomyces fradiae. Appl Microbiol. 1967;15:744–749. doi: 10.1128/am.15.4.744-749.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 7.Mangallam S, Vashi AA, Sukapure RS, Gopalakrishnan KS. Some factors affecting neomycin production byS. fradiae. Hindustan Antibiot Bull. 1974;17:1–6. [PubMed] [Google Scholar]
- 8.Elliah P, Srinivasulu B, Adinarayana K. Optimization studies on neomycin production by a mutant strain ofStreptomyces marinensis in solid state fermentation.Process Biochem. In press.
- 9.Adinarayana K, Ellaiah P, Srinivasulu B, Bhavani Devi R, Adinarayana G. Response surface methodological approach to optimize the nutritional parameters for neomycin production byStreptomyces marinensis under solid state fermentation. Process Biochem. 2003;38:1565–1572. doi: 10.1016/S0032-9592(03)00057-8. [DOI] [Google Scholar]
- 10.Srinivasulu B. Studies on Bioprocess Development With Special Reference to Strain Improvement, Solid State Fermentation and Immobilization ofStreptomyces marinensis Cells for the Production of Neomycin. Visakhapatnam, India: Andhra University; 2002. [Google Scholar]
- 11.Park YS, Ohata N, Okabe M. Neomycin production by partial immobilization ofStreptomyces fradiae on cellulose beads in airlift reactor. J Ferment Bioengg. 1994;78:265–268. doi: 10.1016/0922-338X(94)90303-4. [DOI] [Google Scholar]
- 12.Kierstan M, Bucke C. The immobilization of microbial cells, subcellular organelles, and enzymes in calcium alginate gels. Biotechnol Bioeng. 1977;19:387–397. doi: 10.1002/bit.260190309. [DOI] [PubMed] [Google Scholar]
- 13.Park JK, Chang HN. Microencapsulation of microbial cells. Biotechnol Adv. 2000;18:303–319. doi: 10.1016/S0734-9750(00)00040-9. [DOI] [PubMed] [Google Scholar]
- 14.Farid MA, El Diwany AI, el-Enshasy HA. Production of oxytetracycline and rifamycins B and SV with cells immobilized on glasswool. Acta Biotechnol. 1994;14:67–74. doi: 10.1002/abio.370140112. [DOI] [Google Scholar]
- 15.Adinarayana K, Ellaiah P, Srinivasulu B, Lakshmi Narayana J, Bapi Raju KVVSN. Optimization of medium and cultural conditions for neomycin production using response surface methodology.Ind J Biotechnol. In press.
- 16.Grove DC, Randall WA. Assay Methods of Antibiotics: A Laboratory Manual. New York, NY: Medical Encyclopedia, Inc; 1955. [Google Scholar]
- 17.Ministry for Health and Welfare, Government of India . Indian Pharmacopoeia. New Delhi, India: The Controller of Publications; 1996. pp. A–100. [Google Scholar]
- 18.Mahmoud W, Rehm HJ. Chlortetracycline production with immobilizedStreptomyces aureofaciens. I. Batch culture. Appl Microbiol Biotechnol. 1987;26:333–337. doi: 10.1007/BF00256664. [DOI] [Google Scholar]
- 19.Ellaiah P, Murali Chand G, Srinivasulu B, Pardasaradhi SV. Production of cephalosporin C by immobilized cells ofCephalosporium acremonium. Indian J Exp Biol. 2000;38:1134–1137. [PubMed] [Google Scholar]