• Proceedings of the Korean Society for Applied Microbiology Conference
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• 1986.12a
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• pp.506-508
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• 1986

Metabolic activity of inorganic nitrogenous compounds affects not only microbial growth but also metabolite production in fermentation technology. We have worked on the enzymes participating in ammonia assimulation of some fermentation bacteria. This paper summarizes the results on glutamine synthetase and its application in practical field. Glutamine synthetase (L-glutamate:ammonia ligase, EC. 6.3.1.2) catalyzes the formation of glutamine from glutamate and ammonia at the expense of cleavage of ATP and inorganic phosphate. The enzyme plays a dual role in nitrogen metabolism in bacteria; it is a key enzyme not only in the biosynthesis of various compounds through glutamine but also in the regulation of synthesis of some enzymes involved in the metabolism of nitrogenous compounds. The detailed works with the Eschericia coli and other enterobacterial enzymes revealed that glutamine synthetase is controlled by the following complex of mechanisms: (a) feedback inhibition by end products, (b) repression and derepression of enzyme synthesis, (c) modulation of enzyme activity in response to divalent cation and (d) covalent modification of enzyme protein by adenylylation and its cascade control. Comparative studies have also been made on the enzymes from other organisms.

• Journal of Biosystems Engineering
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• v.41 no.2
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• pp.85-92
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• 2016

Background: The same plant, Camellia sinensis, is used to produce all types of tea, and the differences among the various types arise from the different processing steps that are used. Based on the degree of fermentation, tea can be classified as black, green, white, or oolong tea. Of these, black tea is the most or fully fermented tea. The oxidized polyphenolic compounds such as theaflavins (TF) and thearubigins (TR) formed during fermentation are responsible for the color, taste, flavor, and aroma of black tea. Results: Research indicates that an optimum ratio of TF and TR (1:10) is required to ensure a quality cup of tea. The concentrations of TF and TR as well as desirable quality characteristics increase as fermentation time increases, reaching optimum levels and then degrading if the fermentation time is prolonged. It is also necessary to control the environment for oxidation. There are no established environment conditions that must be maintained during the fermentation of the ruptured tea leaves. However, in most cases, the process is performed at a temperature of $24-29^{\circ}C$ for 2-4 h or 55-110 min for orthodox tea or crush, tear, and curl (CTC) black tea, respectively, under a high relative humidity of 95-98% with an adequate amount of oxygen. Conclusion: The polyphenolic compounds in black tea such as TF and TR as well as un-oxidized catechins are responsible for the health benefits of tea consumption. Tea is rich in natural antioxidant activities and is reported to have great potential for the management of various types of cancers, oral health problems, heart disease and stroke, and diabetes and to have other health benefits such as the ability to detoxify, improve urine and blood flow, stimulate, and improve the immune system.

• Journal of Microbiology and Biotechnology
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• v.16 no.12
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• pp.1849-1855
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• 2006

It has been shown that the initial conditions of bacterial cultivation are extremely important for the successful production of hyaluronic acid (HA) by fermentation. We investigated several parameters that affect cell growth rate and the productivity and molecular weight of hyaluronic acid--i.e., agitation speed, aeration rate, culture temperature, pH, and pressure--to determine how to optimize the production of HA by Streptococcus zooepidemicus on an industrial scale. Using a 30-1 jar fermentor under laboratory conditions, we achieved maximum HA productivity and biomass when the agitation speed and aeration rate were increased simultaneously. By shifting the temperature downward from 35$^{\circ}C$ to 32$^{\circ}C$ at key levels of cell growth during the fermentation process, we were able to obtain HA with a molecular weight of $2.8{\times}10^6$ at a productivity of 5.3 g/l. Moreover, we reproduced these optimized conditions successfully in three 30-1 jar fermentors. By reproducing these conditions in a 3-$m^3$ fermentor, we were able to produce HA with a molecular weight of $2.9{\times}10^6$ at a productivity of 5.4 g/l under large-scale conditions.

• Microbiology and Biotechnology Letters
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• v.24 no.6
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• pp.717-725
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• 1996

We have developed an efficient immobilized cell separator for continuous operation of immobilized fungal cell cultures, and applied this separator to actual fermentation process for the production of cyclosporin A (CyA), a powerful immunosuppressant. In the experiments employing highly viscous polymer (carboxymethyl cellulose) solution, the decantor showed good separating performances at high solution viscosites and fast dilution rates. Air duct and cylindrical separator installed inside the decantor turned out to play key roles for the efficient separation of the immobilized cells. By installing the decantor in an immobilized perfusion reactor system (IPRS), continuous immobilized culture was stably carried out even at high dilution rate for a long period, leading to high productivities of free cells and CyA. Almost no immobilized biomass existed in effuluent stream of the IPRS, demonstrating the effectiveness of the decan- tor system for a long-term continuous fermentation. It was noteworthy that we could obtain these results despite of the unfavorable fermentation conditions, i.e., reduced density of the biosupports caused by overgrowth of cells inside the bead particles and existence of high density of suspended fungal cells (10g/l) in the fermentation broth.

• Archives of Pharmacal Research
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• v.28 no.12
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• pp.1341-1344
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• 2005

Bioassay-guided fractionation of $CHCl_{3}$ extract from the fermentation broth of a sponge Mycale plumose-derived actinomycete Saccharopolyspora sp. nov., led to the isolation of two known prodigiosin analogs - metacycloprodigiosin (1) and undecylprodigiosin (2). These compounds exhibited significant cytotoxic activities against five cancer cell lines: P388, HL60, A-549, BEL­7402, and SPCA4. This is the first report on the significant cytotoxicity of metacycloprodigiosin (1) against human cancer cell lines.

• Asian-Australasian Journal of Animal Sciences
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• v.28 no.10
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• pp.1419-1426
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• 2015

This study aimed to investigate the effect of the ratio of non-fibrous carbohydrates to neutral detergent fibre (NFC/NDF) and undegraded dietary protein (UDP) on rumen fermentation and nitrogen metabolism in lambs. Four $Dorper{\times}thin-tailed$ Han crossbred lambs, averaging $62.3{\pm}1.9kg$ of body weight and 10 mo of age, were randomly assigned to four dietary treatments of combinations of two levels of NFC/NDF (1.0 and 1.7) and two levels of UDP (35% and 50% of crude protein [CP]). Duodenal nutrient flows were measured with dual markers of Yb and Co, and microbial N (MN) synthesis was estimated using $^{15}N$. High UDP decreased organic matter (OM) intake (p = 0.002) and CP intake (p = 0.005). Ruminal pH (p<0.001), ammonia nitrogen ($NH_3-N$; p = 0.008), and total volatile fatty acids (p<0.001) were affected by dietary NFC/NDF. The ruminal concentration of $NH_3-N$ was also affected by UDP (p<0.001). The duodenal flow of total MN (p = 0.007) was greater for lambs fed the high NFC/NDF diet. The amount of metabolisable N increased with increasing dietary NFC:NDF (p = 0.02) or UDP (p = 0.04). In conclusion, the diets with high NFC/NDF (1.7) and UDP (50% of CP) improved metabolisable N supply to lambs.

• Asian-Australasian Journal of Animal Sciences
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• v.30 no.4
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• pp.495-504
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• 2017

Objective: Manipulating the fermentation to improve the performance of the ruminant has attracted the attention of both farmers and animal scientists. Propionate salt supplementation in the diet could disturb the concentration of propionate and total volatile fatty acids in the rumen. This study was conducted to evaluate the effect of calcium propionate supplementation on the ruminal bacterial community composition in finishing bulls. Methods: Eight finishing bulls were randomly assigned to control group (CONT) and calcium propionate supplementation (PROP) feeding group, with four head per group. The control group was fed normal the total mixed ration (TMR) finishing diet, and PROP group was fed TMR supplemented with 200 g/d calcium propionate. At the end of the 51-day feeding trial, all bulls were slaughtered and rumen fluid was collected from each of the animals. Results: Propionate supplementation had no influence the rumen fermentation parameters (p>0.05). Ruminal bacterial community composition was analyzed by sequencing of hypervariable V3 regions of the 16S rRNA gene. The most abundant phyla were the Firmicutes (60.68%) and Bacteroidetes (23.67%), followed by Tenericutes (4.95%) and TM7 (3.39%). The predominant genera included Succiniclasticum (9.43%), Butyrivibrio (3.74%), Ruminococcus (3.46%) and Prevotella (2.86%). Bacterial community composition in the two groups were highly similar, except the abundance of Tenericutes declined along with the calcium propionate supplementation (p = 0.0078). Conclusion: These data suggest that the ruminal bacterial community composition is nearly unchanged by propionate supplementation in finishing bulls.

• Asian-Australasian Journal of Animal Sciences
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• v.22 no.4
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• pp.542-549
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• 2009

An in vitro study was conducted to determine the effect of nitrate-nitrogen used as a sole dietary nitrogen source on ruminal fermentation characteristics and microbial nitrogen (MN) synthesis. Three treatment diets were formulated with different nitrogen sources to contain 13% CP and termed i) nitrate-N diet (NND), ii) urea-N diet (UND), used as negative control, and iii) tryptone-N diet (TND), used as positive control. The results of 24-h incubations showed that nitrate-N disappeared to background concentrations and was not detectable in microbial cells. The NND treatment decreased net $CH_4$ production, but also decreased net $CO_2$ production and increased net $H_2$ production. Total VFA concentration was lower (p<0.05) for NND than TND. Suppression of $CO_2$ production and total VFA concentration may be linked to increased concentration of $H_2$. The MN synthesis was greater (p<0.001) for NND than UND or TND (5.74 vs. 3.31 or 3.34 mg/40 ml, respectively). Nitrate addition diminished methane production as expected, but also increased MN synthesis.

• Asian-Australasian Journal of Animal Sciences
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• v.24 no.4
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• pp.471-478
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• 2011

The objectives were to compare the ability of various rumen microbial fractions to reduce nitrate and to assess the effect of nitrate on in vitro fermentation characteristics. Physical and chemical methods were used to differentiate the rumen microbial population into the following fractions: whole rumen fluid (WRF), protozoa (Pr), bacteria (Ba), and fungi (Fu). The three nitrogen substrate treatments were as follows: no supplemental nitrogen source, nitrate or urea, with the latter two being isonitrogenous additions. The results showed that during 24 h incubation, WRF, Pr and Ba fractions had an ability to reduce nitrate, and the rate of nitrate disappearance for the Pr fraction was similar to the WRF fraction, while the Ba fraction needed an adaptation period of 12 h before rapid nitrate disappearance. The WRF fraction had the greatest methane ($CH_4$) production and the Pr fraction had the greatest prevailing $H_2$ concentration (p<0.05). Compared to the urea treatment, nitrate diminished net gas and $CH_4$ production during incubation (p<0.05), and ammonia-N ($NH_3$-N) concentration (p<0.01). Nitrate also increased acetate, decreased propionate and decreased butyrate molar proportions (p<0.05). The Pr fraction had the highest acetate to propionate ratio (p<0.05). The Pr fraction as well as the Ba fraction appears to have an important role in nitrate reduction. Nitrate did not consistently alter total VFA concentration, but it did shift the VFA profile to higher acetate, lower propionate and lower butyrate molar proportions, consistent with less $CH_4$ production by all microbial fractions.

• Journal of Microbiology and Biotechnology
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• v.20 no.3
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• pp.550-557
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• 2010

Escherichia coli (E. coli) heat-labile enterotoxin B subunit (LTB) was regarded as one of the most powerful mucosal immunoadjuvants eliciting strong immunoresponse to coadministered antigens. In the research, the high-level secretory expression of functional LTB was achieved in P. pastoris through high-density fermentation in a 5-1 fermentor. Meanwhile, the protein was expressed in E. coli by the way of inclusion body, although the gene was cloned from E. coli. Some positive yeast and E. coli transformants were obtained respectively by a series of screenings and identifications. Fusion proteins LTB-6$\times$His could be secreted into the supernatant of the medium after the recombinant P. pastoris was induced by 0.5% (v/v) methanol at $30^{\circ}C$, whereas E. coli transformants expressed target protein in inclusion body after being induced by 1 mM IPTG at $37^{\circ}C$. The expression level increased dramatically to 250-300 mg/l supernatant of fermentation in the former and 80-100 mg/l in the latter. The LTB-6$\times$His were purified to 95% purity by affinity chromatography and characterized by SDS-PAGE and Western blot. Adjuvant activity of target protein was analyzed by binding ability with GMI gangliosides. The MW of LTB-6$\times$His expressed in P. pastoris was greater than that in E. coli, which was equal to the expected 11 kDa, possibly resulted from glycosylation by P. pastoris that would enhance the immunogenicity of co-administered antigens. These data demonstrated that P. pastoris producing heterologous LTB has significant advantages in higher expression level and in adjuvant activity compared with the homologous E. coli system.