• Journal of Microbiology and Biotechnology
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• v.22 no.2
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• pp.207-214
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• 2012

Laccase- and peroxidase-free tyrosinase has commercial importance in the production of L-3, 4-dihydroxyphenylalanine (L-DOPA), which is mainly used in the treatment of Parkinson's disease. In the present study, isolation of an actinomycetes microbial strain capable of producing only tyrosinase is reported. Among all soil isolates, three individual colonies revealed black color around the colony in the presence of tyrosine. Further screening for laccase and peroxidase activities using syringaldazine denoted that one of the isolates, designated as RSP-T1, is laccase and peroxidase negative and produces only tyrosinase. The microbe was authenticated as Streptomyces antibioticus based on 16S ribotyping. Effective growth of this isolate was noticed with the use of medium (pH 5.5) containing casein acid hydrolysate (10.0 g/l), $K_2HPO_4$ (5.0 g/l), $MgSO_4$ (0.25 g/l), L-tyrosine (1.0 g/l), and agar (15 g/l). The scanning electron micrograph depicted that the microbe is highly branched and filamentous in nature. The enzyme production was positively regulated in the presence of copper sulfate. The impact of different fermentation parameters on tyrosinase production depicted that the maximized enzyme titer values were observed when this isolate was grown at 6.5 pH and at $30^{\circ}C$ temperature under agitated conditions (220 rpm). Among all the studied physiological parameters, agitation played a significant role on tyrosinase production. Upon optimization of the parameters, the yield of tyrosinase was improved more than 100% compared with the initial yield.

• Applied Biological Chemistry
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• v.23 no.4
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• pp.211-217
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• 1980

Using Streptomyces chibaensis, which produces a strong inulin-hydrolyzing enzyme, the optimum cultural conditions and composition of the medium for the production of inulase were studied. 1. The highest enzyme activity was obtained at pH 7.5 after 84 hours culture at $30^{\circ}C$. 2. None of carbon source better than inulin was found. 3. $(NH_4)_2HPO_4$ and corn steep liquor were favourable inorganic and organic nitrogen sources for the production of inulase. 4. $KCl,\;MgSO_4\;and\;FeSO_4$ as the metallic salts were effective for the enzyme production at their concentrations of 0.01, 0.05 and 0.0001%, respectively. 5. The highest production of inulase was obtained from the medium of inulin 1.0% and corn steep liquor 2.0% concentrations, respectively.

• Asian-Australasian Journal of Animal Sciences
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• v.30 no.4
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• pp.479-485
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• 2017

Objective: The effects of particle size of processed barley grain, enzyme addition and microwave treatment on in vitro dry matter (DM) disappearance (DMD), gas production and fermentation pH were investigated for feedlot cattle. Methods: Rumen fluid from four fistulated feedlot cattle fed a diet of 860 dry-rolled barley grain, 90 maize silage and 50 supplement g/kg DM was used as inoculum in 3 batch culture in vitro studies. In Experiment 1, dry-rolled barley and barley ground through a 1-, 2-, or 4-mm screen were used to obtain four substrates differing in particle size. In Experiment 2, cellulase enzyme (ENZ) from Acremonium cellulolyticus Y-94 was added to dry-rolled and ground barley (2-mm) at 0, 0.1, 0.5, 1, and 2 mg/g, while Experiment 3 examined the interactions between microwaving (0, 30, and 60 s microwaving) and ENZ addition (0, 1, and 2 mg/g) using dry-rolled barley and 2-mm ground barley. Results: In Experiment 1, decreasing particle size increased DMD and gas production, and decreased fermentation pH (p<0.01). The DMD (g/kg DM) of the dry-rolled barley after 24 h incubation was considerably lower (p<0.05) than that of the ground barley (119.1 dry-rolled barley versus 284.8 for 4-mm, 341.7 for 2-mm; and 358.6 for 1-mm). In Experiment 2, addition of ENZ to dry-rolled barley increased DMD (p<0.01) and tended to increase (p = 0.09) gas production and decreased (p<0.01) fermentation pH, but these variables were not affected by ENZ addition to ground barley. In Experiment 3, there were no interactions between microwaving and ENZ addition after microwaving for any of the variables. Microwaving had minimal effects (except decreased fermentation pH), but consistent with Experiment 2, ENZ addition increased (p<0.01) DMD and gas production, and decreased (p<0.05) fermentation pH of dry-rolled barley, but not ground barley. Conclusion: We conclude that cellulase enzymes can be used to increase the rumen disappearance of barley grain when it is coarsely processed as in the case of dry-rolled barley. However, microwaving of barley grain offered no further improvements in ruminal fermentation of barley grain.

• Journal of Microbiology and Biotechnology
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• v.18 no.9
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• pp.1544-1549
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• 2008

MhMTS and MhMTH are trehalose ($\alpha$-D-glucopyranosyl-[1,1]-$\alpha$-D-glucopyranose) biosynthesis genes of the thermophilic microorganism Metallosphaera hakonensis, and encode a maltooligosyltrehalose synthase (MhMTS) and a maltooligosyltrehalose trehalohydrolase (MhMTH), respectively. In this study, the two genes were fused in-frame in a recombinant DNA, and expressed in Escherichia coli to produce a bifunctional fusion enzyme, MhMTSH. Similar to the two-step reactions with MhMTS and MhMTH, the fusion enzyme catalyzed the sequential reactions on maltopentaose, maltotriosyltrehalose formation, and following hydrolysis, producing trehalose and maltotriose. Optimum conditions for the fusion enzyme-catalyzed trehalose synthesis were around $70^{\circ}C$ and pH 5.0-6.0. The MhMTSH fusion enzyme exhibited a high degree of thermostability, retaining 80% of the activity when pre-incubated at $70^{\circ}C$ for 48 h. The stability was gradually abolished by incubating the fusion enzyme at above $80^{\circ}C$. The MhMTSH fusion enzyme was active on various sizes of maltooligosaccharides, extending its substrate specificity to soluble starch, the most abundant natural source of trehalose production.

• Microbiology and Biotechnology Letters
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• v.19 no.4
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• pp.383-389
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• 1991

In order to produce alkaline protease, psychrotrophic bacterium which have high enzyme activity at low temperature, was isolated by using enrichment culture from various samples and identified as genus alkalopsychrotropic Pseudomonas sp. RP-222. The optimal culture conditions for enzyme production were pH- 10.0, temperature-$20^{\circ}C$ and culture time-4 days. The optimum pH and temperature for the enzyme activity were pH 10.5 and $40^{\circ}C$, respectively and the enzyme was relatively stable at pH 7.0~13.0 and below $50^{\circ}C$. The enzyme was inhibited by ethylenediaminetetraacetate and phenylmethylsulfonylfluoride, indicating that the enzyme was a serine metalloenzyme, but considerably stable in the presence of surface active agents. Activity of the enzyme was increased by the addition of 0.05% Na-$\alpha$-olefin sulfonate.

• Asian-Australasian Journal of Animal Sciences
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• v.15 no.5
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• pp.708-712
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• 2002

Effects of unprocessed (whole) or steam-flaked corn with or without enzyme additives on in vivo nutrient digestibilities and distribution of corn particles in the feces of Holstein steers were determined in a $4{\times}4$ Latin square experiment using four Holstein steers fed the diets containing 1) whole corn without enzyme additive, 2) whole corn with enzyme additive, 3) flaked corn without enzyme additive, or 4) flaked corn with enzyme additive. With regard to nutrient digestibilities such as DM, CP, CF, NFE, NDF, and ADF, no significant differences were detected among treatments, and also the nutrient digestibilities were not affected by the addition of enzyme additive. When distribution of corn particles in the feces was examined, there were no significant differences in the amount of 2, 8 mm and total corn particles. However, feeding flaked corn resulted in less corn particles (4 mm) in the feces than feeding whole corn (p<0.05). There were no significant differences in amounts of corn particles in the feces due to the addition of enzyme additive.

• 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.

• KSBB Journal
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• v.15 no.1
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• pp.88-95
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• 2000

We have produced new-structured oligosaccharides using immoobilized mixed-enzyme reactor of destransucrase from Leuconostoc mesenteroides B-512FMCM and $\alpha$-amylase from Aspergillus oryzae. The reactors of immobilized mixed-exzyme beads were more efficient for the production of oligosaccharides than that of each immobilized enzyme bead in stirred-tank reactior(STR) or in packed-bed reactor(PCR). In continuous flow reactor, the immobilized mixed-enzyme bead in PBR was more stable than in STR, and 52% of initial yield was maintained for 200 hr. New structured-oligosaccharides (NOS) reduced the change of pH in the culture of Streptococcus mutans. It also showed an inhibitory effect on the growth of Staphylococcus aureus.

• Korean Journal of Food Science and Technology
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• v.32 no.2
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• pp.439-443
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• 2000

In order to evaluate the safety of greenhouse horticulture products in Korea, we carried out this work by screening of Fusarium species, which produce deoxynivalenol (DON) from greenhouse horticulture in Western Gyeongnam and Northern Gyeongbuk, Korea. For this study, high sensitive enzyme-linked immunosorbent assay, ALP/NADP method, was applied to detection of DON by enzyme amplification system. From 192 samples of greenhouse horticulture soil and its products, 103 isolates of Fusarium species were obtained. The isolates were cultured at 28C for 15 days and the cultured mediums were extracted by ethyl acetate. The production of DON was verified by thin layer chromatography (TLC). As the results of TLC, 8 strains were identified as DON producing strain. We screened potential producers of DON by ALP/NADP. The levels of DON production were shown from 0.007 to 1.21 g/ml of YES medium. The maximum DON producing strain No. 32-D-3 was isolated from soil in Namhae, Korea. In conclusion, the above results indicate that DON producing fungi contaminated greenhouse horticulture products in Korea. Therefore, further studies are required to accumulate more detailed data about the contamination of DON in various cereals.

• Journal of the Korean Society of Food Culture
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• v.38 no.6
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• pp.442-455
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• 2023

This study investigated the effects of pectinase treatment and skin contact time on the quality characteristics of Dae-hong peach wine. Wine was produced with variations in enzyme treatment and skin contact time (1 hour, 2 hours, 1 day, 2 days, and until the completion of fermentation). Enzyme treatment increased the production yield by 6%, as well as ethanol and redness levels, compared to the non-treated control. Volatile components were higher when the skin contact time was 2 hours or 1 day. Results were compared according to enzyme treatment and skin contact time and found to be influenced by methanol and 3-methyl-1-butanol. Enzyme treatment effectively enhanced yields and volatile compound contents. However, skin contact should be concluded a day before 1 day to ensure compliance with methanol legislative requirements. Therefore, our findings show that enzymatic treatment with shorter skin contact time preserves the distinctive characteristics of Dae-hong peaches and ensures the production of safe and flavorful wine.