• Asian-Australasian Journal of Animal Sciences
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• v.28 no.1
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• pp.50-57
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• 2015

This study was conducted to investigate the effect of soybean meal (SM) and soluble starch (SS) on biogenic amine production and microbial diversity using in vitro ruminal fermentation. Treatments comprised of incubation of 2 g of mixture (expressed as 10 parts) containing different ratios of SM to SS as: 0:0, 10:0, 7:3, 5:5, 3:7, or 0:10. In vitro ruminal fermentation parameters were determined at 0, 12, 24, and 48 h of incubation while the biogenic amine and microbial diversity were determined at 48 h of incubation. Treatment with highest proportion of SM had higher (p<0.05) gas production than those with higher proportions of SS. Samples with higher proportion of SS resulted in lower pH than those with higher proportion of SM after 48 h of incubation. The largest change in $NH_3$-N concentration from 0 to 48 h was observed on all SM while the smallest was observed on exclusive SS. Similarly, exclusive SS had the lowest $NH_3$-N concentration among all groups after 24 h of incubation. Increasing methane ($CH_4$) concentrations were observed with time, and $CH_4$ concentrations were higher (p<0.05) with greater proportions of SM than SS. Balanced proportion of SM and SS had the highest (p<0.05) total volatile fatty acid (TVFA) while propionate was found highest in higher proportion of SS. Moreover, biogenic amine (BA) was higher (p<0.05) in samples containing greater proportions of SM. Histamines, amine index and total amines were highest in exclusive SM followed in sequence mixtures with increasing proportion of SS (and lowered proportion of SM) at 48 h of incubation. Nine dominant bands were identified by denaturing gradient gel electrophoresis (DGGE) and their identity ranged from 87% to 100% which were mostly isolated from rumen and feces. Bands R2 (uncultured bacterium clone RB-5E1) and R4 (uncultured rumen bacterium clone L7A_C10) bands were found in samples with higher proportions of SM while R3 (uncultured Firmicutes bacterium clone NI_52), R7 (Selenomonas sp. MCB2), R8 (Selenomonas ruminantium gene) and R9 (Selenomonas ruminantium strain LongY6) were found in samples with higher proportions of SS. Different feed ratios affect rumen fermentation in terms of pH, $NH_3$-N, $CH_4$, BA, volatile fatty acid and other metabolite concentrations and microbial diversity. Balanced protein and carbohydrate ratios are needed for rumen fermentation.

• Journal of Microbiology and Biotechnology
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• v.27 no.5
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• pp.925-932
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• 2017

Changes in the metabolite profiles of Lactobacillus sakei and its growth media, based on different culture times (0, 6, 12, and 24 h), were investigated using gas chromatography-mass spectrometry (MS) and liquid chromatography-MS with partial least squares discriminant analysis, in order to understand the growth characteristics of this organism. Cell and media samples of L. sakei were significantly separated on PLS-DA score plots. Cell and media metabolites, including sugars, amino acids, and organic acids, were identified as major metabolites contributing to the difference among samples. The alteration of cell and media metabolites during cell growth was strongly associated with energy production. Glucose, fructose, carnitine, tryptophan, and malic acid in the growth media were used as primary energy sources during the initial growth stage, but after the exhaustion of these energy sources, L. sakei could utilize other sources such as trehalose, citric acid, and lysine in the cell. The change in the levels of these energy sources was inversely similar to the energy production, especially ATP. Based on these identified metabolites, the metabolomic pathway associated with energy production through lactic acid fermentation was proposed. Although further studies are required, these results suggest that MS-based metabolomic analysis might be a useful tool for understanding the growth characteristics of L. sakei, the most important bacterium associated with meat and vegetable fermentation, during growth.

• Journal of Food Hygiene and Safety
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• v.17 no.2
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• pp.79-86
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• 2002

Aflatoxin is a secondary fungal metabolite and is a public health hazard because it is a human carcinogenic and has many deleterious effects in men and animals. Rice is one of the better substrates far the fungus which can produce aflatoxins. This study was performed to investigate aflatoxin reduction during the cooking and processing of rice. Aflatoxin was produced by Aspergillus parasiticus ATCC 15517 on well-milled rice (Japonica type) at the level of 13.2 ppb. Cooked rice, rice cakes (baek-sol-gi, plain steamed rice bread), fermented rice (sikhye, sweet rice beverage), and popped rice were prepared from the aflatoxin-contaminated rice. Aflatoxin content in the samples was determined by high performance liquid chromatography. The total aflatoxin level was decreased to 46.9% in the cooked rice, 85.6% in the rice cakes, 11.4% in the fermented rice, and 7.6% in the popped rice, respectively (p.0.05). This reduction brought the level of aflatoxins down to below the Standard and Specification of korea (10 ppb), except for the rice cakes. These results indicate that washing, steaming, fermentation, and popping of rice was helpful in reducing the aflatoxin level in the rice and the most helpful factors were high temperature & high pressure. More research is needed to understand why the preparation of rice cakes did not reduce the level of aflatoxin as much as the other cooking methods.

• Asian-Australasian Journal of Animal Sciences
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• v.32 no.11
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• pp.1705-1714
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• 2019

Objective: Reducing roughage feeding without negatively affecting rumen health is of interest in ruminant nutrition. We investigated the effects of roughage sources and concentrate types on growth performance, ruminal fermentation, and blood metabolite levels in growing cattle. Methods: In this 24-week trial, 24 Hanwoo cattle ($224{\pm}24.7kg$) were fed similar nitrous and energy levels of total mixed ration formulated using two kinds of roughage (timothy hay and ryegrass straw) and two types of concentrate mixes (high starch [HS] and high fiber [HF]). The treatments were arranged in a $2{\times}2$ factorial, consisting of 32% timothy-68% HS, 24% timothy-76% HF, 24% ryegrass-76% HS, and 17% ryegrass-83% HF. Daily feed intakes were measured. Every four weeks, blood were sampled, and body weight was measured before morning feeding. Every eight weeks, rumen fluid was collected using a stomach tube over five consecutive days. Results: The mean dry matter intake (7.33 kg) and average daily gain (1,033 g) did not differ among treatments. However, significant interactions between roughage source and concentrate type were observed for the rumen and blood parameters (p<0.05). Total volatile fatty acid concentration was highest (p<0.05) in timothy-HF-fed calves. With ryegrass as the roughage source, decreasing the roughage inclusion rate increased the molar proportion of propionate and decreased the acetate-to-propionate ratio; the opposite was observed with timothy as the roughage source. Similarly, the effects of concentrate types on plasma total protein, alanine transaminase, Ca, inorganic P, total cholesterol, triglycerides, and creatinine concentrations differed with roughage source (p<0.05). Conclusion: Decreasing the dietary roughage inclusion rate by replacing forage neutral detergent fiber with that from non-roughage fiber source might be a feasible feeding practice in growing cattle. A combination of low-quality roughage with a high fiber concentrate might be economically beneficial.

• Animal Bioscience
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• v.35 no.7
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• pp.999-1009
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• 2022

Objective: To improve the feeding value of rice straw (RS), this study evaluated the potential of rice straw fermented with Pleurotus sajor-caju (FRS) as dairy goat feed. Methods: Spent Pleurotus sajor-caju mushroom substrate was used as fungi inoculum to break the lignocellulose linkage of rice straw, which was solid-fermented at 25℃ to 30℃ for 8 weeks. The ruminal degradation of pangolagrass hay (PG), FRS, and RS were measured in situ for 96 hours in three dry Holstein cows, respectively. Effect of fungi fermented RS on milking performance was studied in feeding trials. A total of 21 Alpine goats a trial were divided into 3 groups: a control group in which PG accounted for 15% of the diet dry matter, and FRS or RS was used to replace the PG in the control group. Goats were fed twice a day under two 28-day trial in individual pens. Meanwhile, a 3×3 Latin square trial (14 days/period) was conducted to study the rumen digestion of three diets by using three fistulated dry goats. Rumen contents were collected for metabolite analyses every one to three hours on the last two days. Results: In situ study showed that fermentation could elevate the rumen degradable fraction and effective degradability of RS (p<0.05). Effective degradability of FRS dry matter was significantly increased from 29.5% of RS to 41.7%. Lactating trial results showed that dry matter intake and milk yield in the PG group and FRS group were similar and higher than those in RS group (p<0.05). The concentration of propionic acid and total volatile fatty acid in the RS group tended to be lower than those in PG group (p<0.10). There were no differences in rumen pH value and ammonia nitrogen level among the groups tested. Conclusion: Fermentation of rice straw by spent Pleurotus sajor-caju mushroom substrate could substantially enhance its feeding value to be equivalent to PG as an effective fiber source for dairy goat. The fermented rice straw is recommended to account for 15% in diet dry matter.

• The Journal of the Convergence on Culture Technology
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• v.8 no.6
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• pp.943-951
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• 2022

In this study, 8 kinds of fruits and vegetables such as apples, pears and radishes were cut and hot water extracts and Steamed hot water extract from fruits and vegetables were prepared and used as experimental substrates. As a result of fermenting with 1% (W/V) red ginseng extract (W/V) and 8 types of lactic acid bacteria mixed starter added to the lactic acid bacteria fermented extract, the pattern and content of ginsenosides were almost unchanged in the fruit and vegetable extract group and the steam treatment group. However, in the lactic acid bacteria fermented group, the TLC pattern was changed according to the fermentation process and treatment, and the content of ginsenosides converted into Rg₃(S) and Rg₅ increased. No change in the number of lactic acid bacteria (cfu) was observed in all four types of fruit and vegetable extracts. The number of lactic acid bacteria CFU was slightly decreased in the four fermented groups of fruit and vegetable extracts, but the growth inhibitory effect of beneficial bacteria was not significant. The growth inhibitory effect of the three harmful bacteria was not affected by the growth of E. coli and Pseudomonas in the four fruit and vegetable extracts. However, the proliferation of Salmonella was inhibited, which was confirmed as the growth inhibitory effect of the fruit and vegetable extract regardless of whether the steamed hot water extract or red ginseng extract was added.

• Journal of Microbiology
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• v.38 no.3
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• pp.160-168
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• 2000

A-factor I a microbial hormone that can positively control cell differentiation leading to spore formation and secondary metabolite formation in Streptomyces griseus. to identify a protease that is deeply involved in the morphological and physiological differentiation of Streptomyces, the proteases produced by Streptomyces griseus IFO 13350 and its A-factor deficient mutant strain, Streptomyces griseus HH1, as well as Streptomyces griseus HH1 transformed with the afsA gene were sturdied. In general Streptomyces griseus showed a higher degree of cell growth and protease activity in proportion to its ability to produce a higher amount of A-factor. In particular, the specific activity of the trypsin of Streptomyces griseus IFO 13350 was greatly enhanced more than twice compared with that of Streptomyces griseus HH1 in the later stage of growth. The specific activity of the metalloprotease of Streptomyces griseus HH1 was greatly enhanced more than twice compared with that of Streptomyces griseus IFO 13350, and this observation was reversed in the presence of thiostreptione, However, Streptomyces griseus HH1 transformed with the afsA gene showed a significantly decreased level of trypsin and metalloprotease activity compared with that of the HH1 strain. There was no significant difference between Streptomyces griseus IFO 13350 and HH1 strain in their chymotrypsin and thiol protease activity, yet the level of leu-amionpeptidase activity was 2 times higher in Streptomyces griseus HH1 than in strain IFO 13350 . Streptomyces griseus HH1 harboring afsA showed a similar level of enzyme activity , however, all the three protease activities sharply increased and the thiol protease activity was critically increased at the end of the fermentation. When a serine protease inhibitor, pefabloc SC, and metalloprotease inhibitor, EDTA, were applied to strain IFO 13350 to examine the in vivo effects of the protease inhibitors on the morpholofical differentiation, the formation of aerial meycelium and spores was delayed by two or three days.

• Journal of Microbiology and Biotechnology
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• v.24 no.2
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• pp.168-176
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• 2014

The morphology of filamentous fungi closely correlates with the productivity in submerged culture. Using itaconic acid (IA) production by Aspergillus terreus as a research model, the quantitative relationship between the growth form of A. terreus and IA production was investigated. IA fermentation was scaled up from shake flasks to a 7 L stirred tank bioreactor based on the quantitative relationship. Our results demonstrated the following: (1) Three morphologies of A. terreus were formed by changing the inoculum level and shape of the flask. (2) Investigation of the effects of the three morphologies on broth rheology and IA production revealed the higher yield of IA on dry cell weight (DCW, IA/DCW) and yield of glucose on DCW (consumed glucose/DCW) were achieved during clump growth of A. terreus. (3) By varying the $KH_2PO_4$ concentration and culture temperature, the relationships between clump diameter and IA production were established, demonstrating that the yield of IA on DCW ($R^2$ = 0.9809) and yield of glucose on DCW ($R^2$ = 0.9421) were closely correlated with clump diameter. The optimum clump diameter range for higher IA production was 0.40-0.50 mm. (4) When the clump diameter was controlled at 0.45 mm by manipulating the mechanical stress in a 7 L fermentor, the yield of IA on DCW and yield of glucose on DCW were increased by 25.1% and 16.3%, respectively. The results presented in this study provide a potential approach for further enhancement of metabolite production by filamentous fungi.

• Food Science and Biotechnology
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• v.18 no.4
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• pp.1046-1050
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• 2009

Our previous study showed that isoflavone profile of soybean undergoes a significant change during cheonggukjang preparation. In particular, the content of metabolite(s) with similar retention time to glycitein under the high performance liquid chromatography (HPLC) condition was significantly increased while the levels of genistein and its derivatives were notably lowered. Therefore, we hypothesized that genistein and its derivatives might be converted to genistein glucosides with similar elution time to glycitein. Our current data suggest that genistein and its derivatives are extensively metabolized into various compounds including genistein glycosides, but not glycitein or its derivatives, by Bacillus species isolated from traditional cheonggukjang. Some of daidzein was also converted into a derivative with shorter retention time by Bacillus amyloliquefaciens 51 and 86-1 but not by Bacillus subtilis 3-5 and 3-17. As metabolism of soy isoflavones, major health-promoting components in soy products, is widely variable depending upon Bacillus species, it is essential to select microorganism that minimizes the breakdown or modification of soy isoflavones in the process of fermented soy product manufacture.

• Journal of Microbiology and Biotechnology
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• v.23 no.5
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• pp.587-601
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• 2013

During the last few decades, it has become evident that the compatibility of the yeast biochemical environment with the ability to process and translate the RNA transcript, along with its capacity to modify a translated protein, are relevant requirements for selecting this host cell for protein expression in several pharmaceutical and clinical applications. In particular, Pichia pastoris is used as an industrial host for recombinant protein and metabolite production, showing a powerful capacity to meet required biomolecular target production levels in high-throughput assays for functional genomics and drug screening. In addition, there is a great advantage to using P. pastoris for protein secretion, even at high molecular weights, since the recovery and purification steps are simplified owing to relatively low levels of endogenous proteins in the extracellular medium. Clearly, no single microexpression system can provide all of the desired properties for human protein production. Moreover, chemical and physical bioprocess parameters, including culture medium formulation, temperature, pH, agitation, aeration rates, induction, and feeding strategies, can highly influence product yield and quality. In order to benefit from the currently available wide range of biosynthesis strategies using P. pastoris, this mini review focuses on the developments and technological fermentation achievements, providing both a comparative and an overall integration analysis. The main aim is to highlight the relevance and versatility of the P. pastoris biosystem to the design of more cost-effective microfactories to meet the increasing demands for recombinant membrane proteins and clinical antibodies for several therapeutic applications.