• Journal of Microbiology and Biotechnology
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• v.11 no.5
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• pp.812-818
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• 2001

Alginate and alginate-derived oligomannuronate enhanced penicillin production in shake flask and fermentor cultures of Penicillium chrysogenum Wis 54-1255 (containing a single copy of the penicillin gene cluster) and in the high producter strain P. chrysogenum AS-P-99 (containing multiple copies of the penicillin gene cluster). Alginate was not used as a single carbon source by P. chryogenum. The stimulatory effect on penicillin production was observed in a defined medium and, to a lower extent, in a complex production medium containing corn steep liquor. Alginate-supplemented cells showed higher transcript levels of the three penicillin biosynthetic genes, pcbAB, pcbC, and penDE, than cells grown in the absence of alginate. The promoters of the pcbAB, pcbC, and penDE genes were coupled to the reporter lacZ gene and introduced as monocopy constructions in P. chrysogenum Wis 54-1225 npe10 by targeted integration in the pyrG locus; the reporter ${\beta}$-galactosidase activity expressed from the three promoters was stimulated by alginate added to the culture medium of the transformants. These results indicate that the stimulation of penicillin production by alginate was derived from an increase in the transcriptional activity of the penicillin biosynthesis genes. The induction by alginate of the transcription of the three penicillin biosynthetic genes is good example of the coordinated induction of secondary metabolism genes by elicitors of plant (or microbial) origin.

• Journal of Microbiology
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• v.34 no.4
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• pp.320-326
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• 1996

To analyze the effects of host versus plasmid on survival of 2, 4-degrading bacteria in environmental samples, strains Pseudomonas cepacia/pJP4, Alcaligenes JMP228/pJP4, P. cepacia/p712, and Alcaligenes JMP228/p712 were separately inoculated into samples of field soil, paddy soil, lake water, and river water, and then the changes of their populations were measured. The strains used contained a 2, 4-D degradative plasmid, either pJP4 conferring fast-growing property to the host or p712 conferring slow-growing property, and were resistant to antibiotics such that the inoculated strains could be enumerated against the indigenous microbial populations. In sterile environmental samples, these strains were stably maintained at the levels used for inoculation, except in sterile paddy soil where Alcaligenes JMP228 strains died drapidly. In natural soil samples for four strains declined steadily with time, but in naturla water samples their polulations fell rapidly at the early phase and then remained almost constant. When the environmentla samples were treated with 2, 4-D, P. cepacia/pJP4 and P. cepacia/p712 maintained significant numbers, while Alcaligenes JMP228/pJP4 and Alcaligenes JMP228/p712 declined significantly in most of the samples. The results indicated that the survivability of genetically modified microorganisms could vary depending on the environments and that their abundance in the environments under s2, 4-D selection was markedly influenced by the nature of the 2, 4-D degradative plasmid as well as type of the host strain.

• Journal of Crop Science and Biotechnology
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• v.10 no.3
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• pp.167-174
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• 2007

Spongy Alphonso mangoes were found to be infected with Staphylococcus bacteria. A Gram positive Staphylococcus strain was isolated from spongy pulp and identified from CABI Bioscience, UK, by partial 16S rDNA sequence analysis and by morphological and biochemical characterization through IMTECH, Chandigarh, India. Although identification by both of these methods indicated the organism belonged to same genus, different species names were given. Changes in total phenolics, reducing, and non-reducing sugars, respiration rate, total carotenoids, peroxidase(POX), and catalase activities were monitored during ripening of these fruits. The climacteric rise in spongy fruits was marked by an increase in respiration rate and a decrease in sugar content. Total phenolics content increased in spongy fruits as compared to ripe non-spongy fruits. Development of corky white tissue in spongy fruits was associated with about a 2.5-fold reduction in total carotenoids and a concomitant increase in lipoxygenase-mediated, $\beta$-carotene co-oxidation. A marked decrease in soluble protein content and about a 1.5-fold increase in POX activity was observed. Maximum POX activity was confined to 50-70%$(NH_4)_2SO_4$ fraction. The intense dark bands visible after POX specific substrate staining of the Native gel indicated a high expression of isoenzymes of POX in spongy fruits. Similarly, changes in levels of catalase activity were also observed in spongy fruits. The results suggest that infection of Alphonso mangoes with Staphylococcus bacteria affects the normal ripening processes of the fruit interfering with the carbohydrate and carotenoid metabolism. Also, the studies indicate the expression of POX and catalase enzymes as a plant defense response to microbial invasion.

• The Plant Pathology Journal
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• v.31 no.2
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• pp.165-175
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• 2015

Anthracnose is a fungal disease caused by Colletotrichum species that is detrimental to numerous plant species. Anthracnose control with fungicides has both human health and environmental safety implications. Despite increasing public concerns, fungicide use will continue in the absence of viable alternatives. There have been relatively less efforts to search antagonistic bacteria from mudflats harboring microbial diversity. A total of 420 bacterial strains were isolated from mudflats near the western sea of South Korea. Five bacterial strains, LB01, LB14, HM03, HM17, and LB15, were characterized as having antifungal properties in the presence of C. acutatum and C. gloeosporioides. The three Bacillus atrophaeus strains, LB14, HM03, and HM17, produced large quantities of chitinase and protease enzymes, whereas the B. amyloliquefaciens strain LB01 produced protease and cellulase enzymes. Two important antagonistic traits, siderophore production and solubilization of insoluble phosphate, were observed in the three B. atrophaeus strains. Analyses of disease suppression revealed that LB14 was most effective for suppressing the incidence of anthracnose symptoms on pepper fruits. LB14 produced antagonistic compounds and suppressed conidial germination of C. acutatum and C. gloeosporioides. The results from the present study will provide a basis for developing a reliable alternative to fungicides for anthracnose control.

• Journal of Life Science
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• v.12 no.2
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• pp.200-207
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• 2002

This study was carried out to isolate of antagonistic bacterium against Pythium ultimum and Rhizoctonia solani AG-4, causal pathogens of vegetables damping-off. Total of 600 strains were isolated from soil and plait roots. The isolates were screened for antagonism against Pythium ultimum and Rhizoctonia solani AG-4. One strain, named YJ-37, was sellected for detained study among those microoganisms screened. It was identified as Bacillus ehimensis based on morphological and physiological characterisitics according to the Bergey's mannual of systematic bacteriology, Sherlock system of Microbial ID Inc. and 16S rDNA sequences methods. Furthermore Bacillus ehimensis YJ-37 showed antifungal activities against Alternaria altrata, Collectotrichum gloeosporioides, Didymella bryoniae, Fusarium moniliforme, Fusarium oxysporum, F. oxysporum cucumerinum, F. oxysporum niveum, Gloeosporium sp., Glomerella sp., G. cingulata, G. lagenaria, Penicillium digitatum, P. italicum, Phytophthora capsici, Sclerotinia sclerotiorum, and Stemprhylium solani.

• The Plant Pathology Journal
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• v.29 no.2
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• pp.193-200
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• 2013

Trichoderma asperellum SKT-1 is a microbial pesticide that is very effective against various diseases. Our study was undertaken to evaluate T. asperellum SKT-1 for induction of resistance against yellow strain of Cucumber mosaic virus (CMV-Y) in Arabidopsis plants. Disease severity was rated at 2 weeks post inoculation (WPI). CMV titre in Arabidopsis leaves was determined by indirect enzyme-linked immunosorbent assay (ELISA) at 2 WPI. Our results demonstrated that among all Arabidopsis plants treated with barley grain inoculum (BGI) of SKT-1 NahG and npr1 plants showed no significant reduction in disease severity and CMV titre as compared with control plants. In contrast, disease severity and CMV titre were significantly reduced in all Arabidopsis plants treated with culture filtrate (CF) of SKT-1 as compared with control plants. RT-PCR results showed increased expression levels of SA-inducible genes, but not JA/ET-inducible genes, in leaves of BGI treated plants. Moreover, expression levels of SA- and JA/ET-inducible genes were increased in leaves of CF treated plants. In conclusion, BGI treatment induced systemic resistance against CMV through SA signaling cascade in Arabidopsis plants. While, treatment with CF of SKT-1 mediated the expression of a majority of the various pathogen related genes, which led to the increased defense mechanism against CMV infection.

• Korean Journal of Environmental Agriculture
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• v.20 no.1
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• pp.57-62
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• 2001

Twelve antagonistic strains against Fusarium wilt of watermelon-rootstock gourd were selected from 54 bacterial isolates which were isolated from the rhizosphere of crop plants growing in various locations. They showed strong inhibitory effects on growth of Fusarium osysporum f. sp. niveum, the causal agent of watermelon-rootstock gourd Fusarium wilt. Among these antagonists, the isolate YJ-3 was the most pronounced in growth-promoting ability for watermelon-rootstock gourd. The growth of watermelon-rootstock gourd in bed soil inoculated with YJ-3 was better by 46 and 13% than those in commercial bed soil alone and in bed soil inoculated with commercial microbial inoculant, respectively. The antagonistic plant growth-promoting rhizobacterium, strain No. YJ-3, was identified as Bacillus sp. on MIDI system. Furthermore, Bacillus sp. YJ-3 showed antifungal activity on growth against Alternaria cucumerina, Botrytis cinerea, Colletotrichum orbiculare, Didymella bryoniae, Rhizoctonia solani and Fusarium oxysporum.

• Microbiology and Biotechnology Letters
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• v.39 no.3
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• pp.287-293
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• 2011

The purpose of this study was to isolate and characterize fungal deteriogens, which induce discoloration of the cement tunnel, and calcite forming bacteria (CFBs), which have antifungal activity against fungal deteriogens. Isolation of mold, bacteria and yeast was performed using several solid media and partially identified using internal transcribed spacer (ITS); 5.8S rRNA gene sequencing and 16s rDNA sequencing. A total of 19 microbial strains were identified with the most widely distributed fungal strain being Cladospirum sphaerospermum. In addition, five bacteria derived from the tunnel were identified as CFBs. Amongst the latter, Bacillus aryabhatti KNUC205 exhibited antifungal activity against Cladospirum sphaerospermum KNUC253 and Aspergillus niger KCTC6906 as concentrated filtered supernatants.

• Journal of Microbiology and Biotechnology
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• v.22 no.6
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• pp.818-825
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• 2012

Keratinases are exciting keratin-degrading enzymes; however, there have been relatively few studies on their immobilization. A keratinolytic protease from Chryseobacterium sp. kr6 was purified and its partial sequence determined using mass spectrometry. No significant homology to other microbial peptides in the NCBI database was observed. Certain parameters for immobilization of the purified keratinase on chitosan beads were investigated. The production of the chitosan beads was optimized using factorial design and surface response techniques. The optimum chitosan bead production for protease immobilization was a 20 g/l chitosan solution in acetic acid [1.5% (v/v)], glutaraldehyde ranging from 34 g to 56 g/l, and an activation time between 6 and 10 h. Under these conditions, above 80% of the enzyme was immobilized on the support. The behavior of the keratinase loading on the chitosan beads surface was well described using the Langmuir model. The maximum capacity of the support ($q_m$) and dissociation constant ($K_d$) were estimated as 58.8 U/g and 0.245 U/ml, respectively. The thermal stability of the immobilized enzyme was also improved around 2-fold, when compared with that of the free enzyme, after 30 min at $65^{\circ}C$. In addition, the activity of the immobilized enzyme remained at 63.4% after it was reused five times. Thus, the immobilized enzyme exhibited an improved thermal stability and remained active after several uses.

• Microbiology and Biotechnology Letters
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• v.45 no.1
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• pp.51-56
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• 2017

Cellulosic biomass is a renewable source for biofuel production from non-edible biomass. An optimized pretreatment process is required for the efficient utilization of cellulosic biomass. Among various pretreatment processes, the use of ionic liquids has been reported recently. However, the residual ionic liquid after pretreatment acts as an inhibitor of microbial fermentation. Recently, we isolated mutant Saccharomyces cerevisiae strains resistant to the ionic liquid 1-ethyl-3-methylimidazolium acetate ([EMIM][Ac]) by using a directed evolutionary approach. When 3% [EMIM][Ac] was added to a medium containing 80 g/l of glucose, mutants D452-B2 and D452-S3 produced 35.6 g/l and 36.3 g/l of ethanol, respectively, for 18 h while the parental strain (S. cerevisiae D452-2) produced 1.3 g/l of ethanol. Thus, these mutant S. cerevisiae strains might prove advantageous when ionic liquids are used for biofuel production from cellulosic biomass.