• Journal of Microbiology and Biotechnology
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• v.32 no.6
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• pp.749-760
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• 2022

α-Galactosidase is a debranching enzyme widely used in the food, feed, paper, and pharmaceuticals industries and plays an important role in hemicellulose degradation. Here, T26, an aerobic bacterial strain with thermostable α-galactosidase activity, was isolated from laboratory-preserved lignocellulolytic microbial consortium TMC7, and identified as Parageobacillus thermoglucosidasius. The α-galactosidase, called T26GAL and derived from the T26 culture supernatant, exhibited a maximum enzyme activity of 0.4976 IU/ml when cultured at 60℃ and 180 rpm for 2 days. Bioinformatics analysis revealed that the α-galactosidase T26GAL belongs to the GH36 family. Subsequently, the pET-26 vector was used for the heterologous expression of the T26 α-galactosidase gene in Escherichia coli BL21 (DE3). The optimum pH for α-galactosidase T26GAL was determined to be 8.0, while the optimum temperature was 60℃. In addition, T26GAL demonstrated a remarkable thermostability with more than 93% enzyme activity, even at a high temperature of 90℃. Furthermore, Ca²⁺ and Mg²⁺ promoted the activity of T26GAL while Zn²⁺ and Cu²⁺ inhibited it. The substrate specificity studies revealed that T26GAL efficiently degraded raffinose, stachyose, and guar gum, but not locust bean gum. This study thus facilitated the discovery of an effective heat-resistant α-galactosidase with potent industrial application. Meanwhile, as part of our research on lignocellulose degradation by a microbial consortium, the present work provides an important basis for encouraging further investigation into this enzyme complex.

• Korean Journal of Agricultural Science
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• v.48 no.1
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• pp.21-31
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• 2021

Yeast strains are capable of hydrolyzing non-digestible saccharides, such as melibiose, raffinose, and stachyose, found in soy meal components. This study revealed the biochemical properties of fermented soybean meal during 72 hours with kefir. Starchyose and raffinose, non-digestible components, were almost digested in kefir 150 mL + soybean meal 500 g + water 70 mL and galactose was produced. Proteolysis of the soybean meal produced most of the small molecule peptides in kefir 150 mL + soybean meal 500 g + water 70 mL. The production of the vitamin B group and C were the highest in kefir 250 mL + soybean meal 500 g. The yeast number of the fermented soybean meal was 7.0 × 10⁶ CFU·mL^-1 which was the highest in kefir 250 mL + soybean meal 500 g. The lactic acid bacteria of the fermented soybean meal was the highest at 3.5 × 10⁹ CFU·mL^-1 in kefir 70 mL + soybean meal 500 g. The antioxidant effect was the highest at 57% in kefir 250 mL + soybean meal 500 g. Expression of inflammation-related cytokine (interleukin [IL]-1β, tumor necrosis factor [TNF]-α, and interleukin [IL]-6) was significantly inhibited in fermented soybean meals with different treatments. These results suggest that fermented soybean meal by kefir has an antiinflammatory and anti-oxidation activity and could be utilized in feed manufacturing, and inhydrolyzing non-digestible soy meal components.

• The Korea Journal of Herbology
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• v.36 no.6
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• pp.47-61
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• 2021

Objectives : 'Steaming and drying' is a traditional processing method that has been used to produce Suk-ji-hwang (熟地黃; Rehmanniae Radix Preparata) from Ji-hwang (地黃, the fresh root of Rehmannia glutinosa Liboschitz ex Steudel; Rehmanniae Radix). The steaming and drying process, which is proceeded in heating and moisturizing conditions, plays a crucial role in the change of therapeutic effect of Ji-hwang, presumably due to the modification of its chemical constituents. In this article, the chemical influence of the 'Steaming and drying' process was investigated for understanding the underlying mechanism of chemical modification of Ji-hwang. Methods : The articles regarding the modifications of chemical constituents of Ji-hwang during the 'Steaming and drying' process were collected and analyzed to investigate the influence of the processing to Ji-hwang. Results : The results indicated that iridoid glycosides were degraded to their aglycones and sugars, and such degradations occurred faster at a high pressure than at an atmospheric pressure during the process. The contents of catalpol, ajugol, and acteoside were decreased, while those of rehmannioside A and D were slightly increased during the repeated processing. The contents of oligosaccharides, namely sucrose, maltose, raffinose, and stachyose (except for manninotriose), were decreased, while those of monosaccharides, glucose and fructose, were increased by the repeated processing. Conclusions : These results demonstrate that the 'Steaming and drying' process influenced the chemical constituents of Ji-hwang and provide probable basis for the therapeutic modification of Suk-ji-hwang after the processing of Ji-hwang.

• Animal Bioscience
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• v.36 no.10
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• pp.1568-1577
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• 2023

Objective: A study was conducted to determine the chemical composition of banana meal and rice bran from Australia or South-East Asia and test the hypothesis that there are no differences in rice bran produced in different countries, but there are differences between full-fat and defatted rice bran. Methods: Two sources of banana meal and 22 sources of rice bran (full-fat or defatted) from Australia or South-East Asia were used. All samples were analyzed for dry matter, gross energy, nitrogen, amino acids (AA), acid hydrolyzed ether extract (AEE), ash, minerals, total starch, insoluble dietary fiber, and soluble dietary fiber. Banana meal was also analyzed for sugars including glucose, fructose, maltose, sucrose, stachyose, and raffinose. Results: Chemical analysis demonstrated that banana meal from the Philippines is primarily composed of starch. Full-fat rice bran from Australia had greater (p<0.05) concentrations of AEE, lysine, and glycine than samples from the Philippines and Vietnam. Full-fat rice bran from Australia and Thailand had greater (p<0.05) concentrations of gross energy and most AA than rice bran from Vietnam. Full-fat rice bran from Australia had greater (p<0.05) concentrations of tryptophan and manganese than all other sources, but full-fat rice bran from the Philippines contained less (p<0.05) zinc than all other sources of rice bran. Gross energy, AEE, and copper were greater (p<0.05) in full-fat rice bran compared with defatted rice bran, but defatted rice bran contained more (p<0.05) crude protein, ash, insoluble dietary fiber, total dietary fiber, AA, and some minerals than full-fat rice bran. Conclusion: Banana meal is a high-energy source that can be used as an alternative ingredient in livestock diets. Full-fat rice bran from Australia and Thailand contained more concentrations of AEE and AA than samples from the Philippines or Vietnam. Full-fat rice bran had more gross energy and AEE than defatted rice bran, whereas defatted rice bran contained more crude protein, ash, and total dietary fiber.

• Food Science of Animal Resources
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• v.25 no.4
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• pp.483-493
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• 2005

This experiment was carried out to examine the fermentation properties of yogurt prepared with bovine milk and soybean milk at the mixed ratios of 3:1, 2:1, 1:1, 1:2 and 1:3. The effect of bovine milk and soybean milk on promoting the fermentation was higher un pH was $3.75\~4.16$ when Lactobacillus salivarius ssp. salivarius CNU27, lactic culture 1(Lactobacillus delbrueckii ssp. bulgaricus(LB12)), Streptococcus salivarius ssp. thermophilus (ST36) and Lactobacillus acidophilus KCTC3150 were used. Titratable acidity was the highest when lactic culture 1[Lactobacillus delbrueckii ssp. bulgaricus(LB12), Streptococcus salivarius ssp. thermophilus(ST36)] was mea and the mixed ratio of bovine milk and soybean milk was 2:1. The average viable counts of lactic acid bacteria was the highest level of $2.17\times10^9$ cfu/ml when Lactobacillus salivarius ssp. salivarius CNU27 was used, and the mixed ratio of bovine milk and soybean milk was 1:3. the highest lactic acid production was 412.52mM when lactic culture 1[Lactobacillus delbrueckii ssp. bulgaricus (LB12), Streptococcus salivarius ssp. thermophilus (ST36)] was used, and the mixed ratio of bovine milk and soybean milk was 1:1. The production of acetic acid was the highest and the concentration was 394.01mM when lactic culture 2(Bifidobacterium longum, Lactobacillus acidophilus, Streptococcus salivarius ssp. thermophilus) was used and the mixed ratio of bovine milk and soy bean milk was 3:1. Tn the carbohydrate hydrolysis, stachyose was hydrolyzed $53.92\%$ as compared with the control when Lactobacillus salivarius subsp salivarius CNU27 was used, and the mixed ratio of bovine milk and soy bean milk was 1:3. The highest viscosity of yogurt was $1,300\~1,660$ cP when the mixed ratio of bovine milk and soybean milk was 1:3. The overall acceptability, $4.17\pm0.69$, was the highest when Lactobacillus acidophilus KCTC3150 was used and when the mixed ratio of bovine milk and soybean milk was 2:1.

• Journal of the Korean Society of Food Science and Nutrition
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• v.42 no.8
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• pp.1255-1262
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• 2013

The objective of this study was to analyze the general and functional components of various soybeans, used for producing soy sauce, tofu, bean sprouts, and for cooking with rice. The moisture, crude protein, crude fat, and ash content of soybeans were in the ranges of 5.50~6.16%, 38.49~41.08%, 14.89~21.89%, and 4.89~5.86%, respectively. Mineral and functional components varied by sample and showed no relationship with usage type. Somyeong for bean sprouts had the highest level of Ca (406.36 mg/100 g), and Sinpaldal for soy sauce and tofu had the highest level of Mg (247.79 mg/100 g). However, Jinpum No. 2 for soy sauce and tofu had the lowest level of Ca (199.51 mg/100 g), and Seonhuk had the lowest level of Mg (90.03 mg/100 g) among the soybeans tested. The isoflavone content in soybeans was in the range of 97.54~402.00 mg/100 g. Somyeong for bean sprouts had the highest level and Seonheuk for cooking with rice had the lowest level. Total oligosaccharides were in the range of $5,838.52{\sim}9,345.90{\mu}g/100g$. Sucrose content was approximately 50% of total oligosaccharides in all samples. Raffinose content was $516.28{\sim}806.95{\mu}g/100g$, and stachyose content was $2,047.13{\sim}3,454.10{\mu}g/100g$. Phytosterols including bracassisterol, campesterol, stigmasterol, and ${\beta}$-sitosterol ranged from 19.25~35.34 mg/100 g. ${\beta}$-sitosterol represented 50% of total phytosterols in almost all samples, followed by campesterol, stigmasterol and bracassisterol. The phytic acid content in soybeans tested was around 2%, except for Sinpaldal No. 2 and Jinpum No. 2 at 0.86% and 1.65%, respectively. The dietary fiber of soybeans was in the ranges of 24.20%~29.20%.

• Applied Biological Chemistry
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• v.43 no.2
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• pp.86-94
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• 2000

In order to develop the enzymatic hydrolysis system concerned with taste and flavor, strains having the high hydrolyzing activity on the soy protein were selected from some traditional Mejus. Two molds and one bacterium producing enzymes which were different in character of hydrolysis were isolated and identified. Leucine and azodye enzyme activities of both M4 and M5 were relatively high among in the isolated molds. And, leucine enzyme activity of B16 was the lowest in the isolated bacteria. These strains were isolated as microorganisms having a dissimilar hydrolysis pattern on the soy protein by enzymatic reactions. Mold M4 on the culture solid media was mycelium colors of white and its sclerotia colors were changed from white to black. According to the result of slide culture, radial conidial head, subclavate vesicle, conidia of subglobose, stipes of uncolored with smooth walls and metula and phialides were existed. Because M4 was taxonomically similar to the characteristics of Aspergillus oryzae (ahlburg) species, M4 was identified and named as Aspergillus oryzae M4.Mold M5 showed white and black mycelium on the MEA medium. Mold M5 colony exhibited grayish-green color and have long(7 mm) sporangiophores at slide culture. Sporangia became brownish-gray and the wall of larger sporangia was broken to form small collars, and smaller sporangia were fomed continually from large basal membrane. Columella is globose and hyaline, and sporangiospores are ellipsoidal of small diameter$(80\;{\mu}m)$. Because M5 was taxonomically similar to the Mucor circinelloides of zygomycetes, M5 was was identified and named as Mucor circinelloides M5. Bacteria B16 colony was opaque white, circular and lobate, and had rod shaped endospore. B16 was found positive in stain, catalase, ${\beta}-glucosidse$ and V-P tests. B16 was found to utilize D-fructose, ${\alpha}-D-glucose$, maltose, D-mannose, D-raffinose, stachyose and sucrose. By the morphological and physiological results, the characteristics of B16 was thought to correspond to that of Bacillus megaterium. However, fatty acid composition was similar to Paenibacillus marcerans, requiring further study for the definite identification. Accordingly, Bacteria B16 was provisionally classified and named as Bacillus megaterium B16.

• Korean Journal of Food Science and Technology
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• v.34 no.6
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• pp.962-968
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• 2002

Effects of gamma-irradiation of 1 to 10 kGy on the microbial growth, contents of amino acids, fatty acids, and free sugars, and changes in acid values in soybean powder were studied. Irradiation doses at $3{\sim}5\;kGy$ inhibited the mold growth completely in two kinds of imported soybean powders. Contents of sulfur-containing amino acids, such as cysteine, in both soybean powders decreased with irradiation, whereas no significant changes in free amino acid and fatty acid contents of both soybean powders were observed. Free sugar contents of stachyose and sucrose in Chinese soybean powder decreased with increasing irradiation dose level, whereas, those of other sugars remained unchanged. Results of this study confirm that $3{\sim}5\;kGy$ irradiation can be safely applied to apply to soybean powder without causing significant quality deteriorations microbiologically and chemically.

• Microbiology and Biotechnology Letters
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• v.43 no.3
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• pp.195-203
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• 2015

Two bacterial strains, Bacillus licheniformis YB-1413 and YB-1414, producing extracellular α-galactosidase, were obtained from homemade Doenjang. On the basis of their biochemical properties, 16S rRNA sequences and random amplified polymorphic DNA patterns by polymerase chain reaction, they were found to be somewhat different from one another. α-Galactosidase productivities of the two isolates were increased by wheat bran, but drastically decreased by melibiose, raffinose and sucrose which were used as carbon sources. The enzyme productivities were increased by yeast extract as a nitrogen source with maximum levels of 1.87 U/ml for YB-1413 and 1.69 U/ml for YB-1414, respectively. The enzymes of both isolates exhibited maximum activity for hydrolysis of para-nitrophenyl-α-D-galactopyranoside (pNP-αGal) under reaction conditions of pH 6.0 and 45℃. Their hydrolyzing activities for pNP-αGal were drastically decreased by the addition of low concentrations of ribose and galactose. They were capable of hydrolyzing completely α-1,6 linked galactosyl residue in melibiose, raffinose and stachyose, which are known to be anti-nutritional factors in products of soybean and legume. In relation to the latter, the isolates YB-1413 and YB-1414 have potential applicability in improving soybean-fermented foods and the nutritional value of soybean feed.

• Journal of Microbiology and Biotechnology
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• v.19 no.11
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• pp.1295-1300
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• 2009

A 2,172-bp full-length gene (aga-F78), encoding a protease-resistant $\alpha$-galactosidase, was cloned from Rhizopus sp. F78 and expressed in Escherichia coli. The deduced amino acid sequence shared highest identity (45.0%) with an $\alpha$-galactosidase of glycoside hydrolase family 36 from Absidia corymbifera. After one-step purification with a Ni-NTA chelating column, the recombinant Aga-F78 migrated as a single band of ~82 and ~210 kDa on SDS-PAGE and nondenaturing gradient PAGE, respectively, indicating that the native structure of the recombinant Aga-F78 was a trimer. Exhibiting the similar properties as the authentic protein, purified recombinant Aga-F78 was optimally active at $50^{\circ}C$ and pH 4.8, highly pH stable over the pH range 5.0-10.0, more resistant to some cations and proteases, and had wide substrate specificity (pNPG, melidiose, raffinose, and stachyose). The recombinant enzyme also showed good hydrolytic ability to soybean meal, releasing galactose of $415.58\;{\mu}g/g$ soybean meal. When combined with trypsin, the enzyme retained over 90% degradability to soybean meal. These favorable properties make Aga-F78 a potential candidate for applications in the food and feed industries.