• Journal of Microbiology and Biotechnology
• /
• v.33 no.6
• /
• pp.831-839
• /
• 2023

Tylosin is a potent veterinary macrolide antibiotic produced by the fermentation of Streptomyces fradiae; however, it is necessary to modify S. fradiae strains to improve tylosin production. In this study, we established a high-throughput, 24-well plate screening method for identifying S. fradiae strains that produce increased yields of tylosin. Additionally, we constructed mutant libraries of S. fradiae via ultraviolet (UV) irradiation and/or sodium nitrite mutagenesis. A primary screening of the libraries in 24-well plates and UV spectrophotometry identified S. fradiae mutants producing increased yields of tylosin. Mutants with tylosin yield 10% higher than the wild-type strain were inoculated into shake flasks, and the tylosin concentrations produced were determined by high-performance liquid chromatography (HPLC). Joint (UV irradiation and sodium nitrite) mutagenesis resulted in higher yields of mutants with enhanced tylosin production. Finally, 10 mutants showing higher tylosin yield were re-screened in shake flasks. The yield of tylosin A by strains UN-C183 (6767.64 ± 82.43 ㎍/ml) and UN-C137 (6889.72 ± 70.25 ㎍/ml) was significantly higher than that of the wild-type strain (6617.99 ± 22.67 ㎍/ml). These mutant strains will form the basis for further strain breeding in tylosin production.

• 한국생물공학회:학술대회논문집
• /
• 2001.11a
• /
• pp.89-92
• /
• 2001

Aspergillus niger has been used as a host system to express many heterologous proteins. It has various advantages over other expression systems in that it is a small eukaryotic GRAS (Generally Recognized aS Safe) organism with a capacity of secreting large amount of foreign proteins. However, it has been known that the presence of an abundant protease is a limiting factor to express a heterologous protein. The proteases deficient mutants of A. niger were obtained using UV -mutagenesis. A total of 1 ${\times}$ $10^5$ spores were irradiated with 10-20% survival dose of UV, 600J/M2 at 280nm, and the resulting spores were screened on the casein -gelatin plates. Ten putative protease deficient mutants were further analyzed on the starch plates to differentiate the pro from the secretory mutant. An endogenous extracellular enzyme, glucose oxidase, was also examined to confirm that the mutant phenotype was due to the proteases deficiency rather than the mutation in the secretory pathway. The reduced proteolytic activity was measured using SDS-fibrin zymography gel, casein degradation assay, and bio-activity of a supplemented hGM -CSF (human Granulocyte-Macrophage Colony Stimulating Factor). Comparing with the wild type strain, less than 30 % of proteolytic activity was observed in the culture filtrate of the protease deficient mutant (pro -20) without any notable changes in cell growth and secretion.

• Microbiology and Biotechnology Letters
• /
• v.22 no.6
• /
• pp.584-592
• /
• 1994

Several mutation procedures have been compared to obtain an ethanol-tolerant Saccha- romyces diastaticus strain secreting glucoamylase. These procedures include spontaneous mutation, EMS treatment, UV irradiation, and combination of EMS treatment and UV irradiation. All these methods were followed by adaptation of the yeast cells to gradually higher ethanol concentration. Among these procedures, the combined method of EMS treatment and UV irradiation gave the promising result, i.e. the ethanol tolerance of the yeast increased from 11.5%(v/v) to 14.0%(v/v). Respiratory deficient petite mutants of industrial and ethanol-tolerant yeast strains have been isola- ted and hybridized with haploid S. diastaticus strains. The resulting hybrids showed increased ethanol tolerance and starch-fermentability.

• Microbiology and Biotechnology Letters
• /
• v.13 no.1
• /
• pp.41-49
• /
• 1985

Bacillus sphaericus was mutagenized with UV light irradiation and dimethyl sulfate. Thirty-five conditional lethal temperature-sensitive(ts) mutants were isolated at the nonpermissive temperature of $42^{\circ}C$ and classified into three groups by their growth characteristics on the nutrient broth, peptone glucose yeast extract agar and mineral salts agar. First was the lethal ts group, 24 mutants, which did not grow at the nonpermissive temperature, the second, 9 mutants, was the less growth is group whose growth was restricted to one-half, and the third, 2 mutants, was the cold lethal ts group whose growth was restricted at the permissive temperature($25^{\circ}C$and $30^{\circ}C$)

• Journal of Microbiology and Biotechnology
• /
• v.26 no.5
• /
• pp.854-866
• /
• 2016

The production cost of biodiesel from microalgae is still not competitive, compared with that of petroleum fuels. The genetic improvement of microalgal strains to increase triacylglycerol (TAG) accumulation is one way to reduce production costs. One of the most promising approaches is the isolation of starch-deficient mutants, which have been reported to successfully increase TAG yields. To date, such a stable mutant is not available in an oleaginous marine microalga, despite several advantages of using marine species for biodiesel production. Algae in the genus Dunaliella are known to tolerate high salt concentration and other environmental stresses. In addition, the cultivation processes for large-scale outdoor commercialization have been well established for this genus. In this study, Dunaliella tertiolecta was used to screen for starch-deficient mutants, using an iodine vapor-staining method. Four out of 20,016 UV-mutagenized strains showed a substantial reduction of starch content. A significantly higher TAG content, up to 3-fold of the wild-type level, was observed in three of the mutants upon induction by nitrogen depletion. The carotenoid production and growth characteristics of these mutants, under both normal and oxidative stress conditions, were not compromised, suggesting that these processes are not necessarily affected by starch deficiency. The results from this work open up new possibilities for exploring Dunaliella for biodiesel production.

• Applied Chemistry for Engineering
• /
• v.24 no.2
• /
• pp.155-160
• /
• 2013

Research on mutant generation and isolation for microalgae yielding enhanced lipid accumulation is an important issue for the production of economic biodiesel. In the present study, ultraviolet (UV-B type) ray induced mutant generation was tried using a photosynthetic microalgae, Nannochloropsis oculata (N. oculata), for the production of biodiesel. The resulting colonies were isolated and further cultured with both liquid and solid state f/2 media. After a few week cultivation, changes of cell growth rate, dry cell weight, and several important intracellular components (chlorophyll, carotenoid, and lipid) were investigated. Two mutants among thousands colonies showed an increased cell growth and high lipid accumulation as compared to those of wild type. It was also observed that the increased cell growth rate is associated with the overexpressed intracellular proteins. However, the mutants showed a decrease in the chlorophyll biosynthesis.

• Mycobiology
• /
• v.32 no.2
• /
• pp.88-94
• /
• 2004

Protoplasts of the wild type strain of Pleurotus osteatus were mutagenized with UV light, and 3,000 colonies were examined for abnormal mycelial and fruiting phenotypes. Forty one strains displayed variant phenotypes in mycelia and fruiting processes. The variant phenotypes were classified into 6 groups: (1) auxotrophic strains, which are incapable of growing on minimal media and can only grow when provided with their specific requirements; (2) abnormal vegetative strains, which grow very slowly on minimal and complete media; (3) primordiumless strains, which fail to develop to the formation of primordia; (4) maturationless strains, which form primordia, but do not form mature fruiting bodies; (5) specifically colored strains, which have Specific bluish grey or bluish white pileus; (6) poorly spored strains, which fail to produce basidiospore or which produce few spores. These variant strains may be useful in genetic breeding programs and for the studies of fungal development and genetics.

• The Korean Journal of Mycology
• /
• v.13 no.2
• /
• pp.75-78
• /
• 1985

Mutagenesis of protoplast could serve a great potential tool for improvement of strains and genetics in higher fungi. For the isolation of auxotrophic mutants from protoplasts of Pleurotus ostreatus and Pleurotus florida, viability levels of ultraviolet lights were determined. Seven auxotrophs were obtained from protoplasts irradiating UV to give $0.83{\sim}15%$ survival. The mutants showed a single requirement for each of Arg, Ribo-l, Ribo-2 or Phen for growth. Some of them showed two or three kinds of requirements, Gly Ser, Ade Hypo or Ala Om Tryp for growth.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.22 no.5
• /
• pp.636-642
• /
• 1993

To improve the quality of mirin, various molds were screened for mutants with high acid carboxypeptidase (ACPase) by the method of ultraviolet radiation. Mutants, Aspergillus oryzae 9-12 and Aspergillus shiroussamii 6082-60 showed activities of ACPase about 2~6 times higher than their parent strains. Aspergillus oryzae 9-12 and Aspergillus shirosamii 6082-60 were the most suitable strains for preparing koji in mirin by the conventional or improved methods. The results showed that total sugar, reducing sugar and total nitrogen were almost the same values in mirin prepared by both methods. The yield of mirin was higher in the improved method than in the conventional method. The clouding formation of mirin appeared in the conventional method ; however, mirin prepared with the mutant koji by the improved method did not show clouding formation.

• Applied Biological Chemistry
• /
• v.18 no.1
• /
• pp.16-22
• /
• 1975

This experiment was carried out to investigate the physiological characteristics of two original yeasts, 5-Y-5 and 6-Y-6, which selected from 24 Takju yeasts and three mutants, 30-24,30-81 and 40-27. induced from two original yeasts by the irradiation of UV light. The results were summarized as follows. 1) Alcohol tolerances of three mutants were decreased in some degree as compared with those of original yeasts. 2) Tolerances of lactic and citric acids of acid producing mutant 30-81, was increased than those of original yeasts. 3) In the case of using ammonium sulfate as a nitrogen source, two original yeasts and three mutants required Ca-pantothenate as a essential growth factor and four strains of yeasts except the mutant, 30-81, required biotin as a stimulated growth factor, When asparagine was used as a nitrogen source, two original yeasts and three mutants showed the same as above result but the stimulated effect of biotin was far less. 4) Propagation powers of the mutants were weaken than those of original yeasts, particular that of acid producing mutant, 30-81, was the weakest in the three mutants. 5) The optimum temperature for fermentation of original yeasts were $30^{\circ}C\;to\;35^{\circ}C$ but three mutants were $25^{\circ}C\;to\;30^{\circ}C$. 6) The optimum pH for fermentation of original yeasts were pH 5 to 6, and there is no appreciable difference between original yeasts and three mutants. The fermentation power of mutant,30-81, was decreased more rapidly than those of other mutants according to approach neutral. Three mutants were more sensible to heat than original yeasts. 7) Two original yeasts and three mutants were inhibited more over 20 percent of sugar for fermentation and three mutants were more sensible to sugar concentration than original yeasts.