• Title/Summary/Keyword: Raffinose

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Isolation and Identification of Microorganisms Producing the Soy Protein-Hydrolyzing Enzyme from Traditional Mejus (전통메주로부터 대두단백질 가수분해효소 생산성 미생물의 분리 및 동정)

  • Kang, Min-Jung;Kim, Seong-Ho;Joo, Hyun-Kyu;Lee, Gap-Sang;Yim, Moo-Hyun
    • 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.

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Cryopreservation of CHO Cell using Serum-Free Media (무혈청 배지를 이용한 CHO 세포의 동결보존)

  • Kim, Yoo-Kang;Park, Hong-Woo;Choe, Tae-Boo
    • KSBB Journal
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    • v.21 no.2
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    • pp.110-117
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    • 2006
  • During routine maintenance, animal cell lines are commonly cryopreserved in growth medium containing serum with 10% DMSO. But, in case of bioprocess under the serum-free conditions, including cultivation of cell lines and producing of pharmaceuticals, the cryopreservation should be executed without serum to prevent a cross-contamination. This experiments were performed to investigate the effects of the serum-free cryopreservation on the CHO cells. To improve the survival rates of the cryopreserved CHO cells in serum-free condition, first, the effects of permeable and non-permeable additives for substitute serum on cell viability were investigated. The combination of 10% DMSO and 0.03 M raffinose in MEM-${\alpha}$ without serum indicated 76% of cell viability. However, it did not reach the survival rates(more than 95%) of the conventional cryopreservation. In the second, to evaluate the cryopreservative ability of the serum-free medium(SFM) we compared viability of the CHO cells cryopreserved in the SFMs(Sigma C5467, C4726, and C1707, JBI SF486 and PF486), the cryoprotectant(Genenmed CAN-1000) and the MEM-${\alpha}$ with serum. All solution contained 10% DMSO. As a result of the comparison, cryopreserved cells in the SFMs showed over 95% of viability and appeared predominant viability better than cryoprotectant CAN-1000. Finally, we assessed the stability of the CHO cells in the long-term cryopreservation(LTC) using SFM. Every three months, the cryopreserved CHO cells were thawed to estimate the cell viability and the recovery rates. Then, real-time RT-PCR analyzed the inserted CHO DHFR gene. All results for the LTC appeared the same stability as the serum containing cryopreservation. In the conclusion, it could be seen that the LTC in the SFM can substitute for serum using methods in the bioprocess proceeded by CHO cells for more than 18 months.

Studies on Freezing Tolerance of Mulberry, Morus species -Relation between Freezing Tolerance and Some Substances in Mulberry Branches- (뽕나무 내한성에 관한 연구 -특히 지조함유물질과 내한성과의 관계에 대하여-)

  • 김호락
    • Journal of Sericultural and Entomological Science
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    • v.22 no.1
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    • pp.7-25
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    • 1980
  • Some substances and freezing tolerance in the mulberry (Morus species) branch have been studied on the basis of varietal differences and harvesting times along with harvesting methods in autumn. The results obtained are summarized as follows; 1. The highest freezing tolerance was shown in the varieties of Yongcheon-chou, Jasan, Kang-weon No. 3 and Ichihei, the medium in Roso. Kairyonezumigaeshi, Yanagida and Kokuso No. 28, and the lowest in Ichinose, Mokuso, Kokuso No. 21 and Suweousang No. 3. 2. There was a signifiant negative correlation (r= -0.59*) between death atop percentage in the field and the temperature required to kill 50% of the mulberry buds (T$_{50}$) with the harvesting times and methods in autumn. Cold hardening occurred in the early through the end of September with the peak at the mid-september. During this period, leaf harvest decreased freezing tolerance with remarkable decrease due to picking all the leaves and leaving several leaves at the base of branch. Greater cold hardening was induced by leaving several leaves after topping. 3. Negative correlations were observed between freezing tolerance and the contents of soluble (r =-0.70*) and crude (r= -0.70*) protein. However, positive correlations were shown between freezing tolerance and total carbohydrate contents per crude (r=0.31*) and per soluble (r=0.71*) protein . There were also positive correlations between freezing tolerance and total sugar (r=0.67*) and RNA content (r=0.99**). No relationships of dry matter. fat. total carbohydrate and DNA contents were observed to the freezing tolerance. 4. Such sugars as raffinose. lactose, sucrose, glucose, fructose. arabinose. xylose. ribose (assumed) and rhamnose were detected in winter mulberry branch. Major sugars such as sucrose, glucose, and fructose were supposed to have higher relationship to the freezing tolerance than the other sugars. 5. Late harvesting increased RNA content except in the case of total leaf picking at mid-September. Leaf picking decreased RNA content. Some amount of RNA was, however, maintained by leaving several leaves after topping Leaving upper-middle leaves of a branch showed high RNA content. Leaving young leaves at the top and the overmatured leaves at the base showed low content. A positive correlation (r=0.51*) was noted between RNA content and freezing tolerance in the different harvesting methods.s.

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Isolation and Identification of Lactic Acid Bacteria with Probiotic Activities from Kimchi and Their Fermentation Properties in Milk (전통 김치로부터 Probiotic 유산균의 분리 및 우유 발효 특성)

  • Lim, Young-Soon;Kim, JiYoun;Kang, HyeonCheol
    • Journal of Dairy Science and Biotechnology
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    • v.37 no.2
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    • pp.115-128
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    • 2019
  • Lactic acid bacteria obtained from traditional Kimchi were selected on the basis of their caseinolytic activity and lactose usability and examined for availability as a starter in probiotic activity. Thirty-two strains were selected as lactic acid producing bacteria in BCP agar, and two strains (KC23 and KF26) with more than 90% resistance for both acid and bile salts were selected. The two strains were identified as L. plantarum (KC23) and L. paracasei (KF26) by API 50 CHL system and 16S rRNA sequence analysis. L. plantarum (KC23) was finally selected based on its biochemical characteristics for lactose and raffinose usability. Free tyrosine content increased rapidly in 10% skimmed milk medium, from $24.1{\mu}g/mL$ after 8 h to $43.9{\mu}g/mL$ after 16 h. Additionally, the caseinolytic clear zone of 12 mm of L. plantarum (KC23) was greater than the 9 mm zone of commercial L. acidophilus CSLA. The bacterium exhibited mesophilic growth and yielded $8.9{\times}10^8CFU/mL$ when incubated at $37^{\circ}C$ for 12 h at pH 4.25. Moreover, L. plantarum KC23 exhibited antibacterial activity as it formed a clear zone of 8-13 mm for the 5 pathogens. Adherent activity was 2.23 fold higher than that of LGG. The acidity of 10% skimmed milk fermented for 12 h was 0.74%.

Optimal conditions and effects of prebiotics for growth and antimicrobial substances production of Lactobacillus brevis BK11 (Lactobacillus brevis BK11의 증식과 항균물질 생산을 위한 최적 배양조건 및 prebiotics의 영향)

  • Lim, Eun-Seo
    • Korean Journal of Microbiology
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    • v.51 no.3
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    • pp.288-299
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    • 2015
  • Lactobacillus brevis BK11 obtained from Baikkimchi was selected to study the effects of culture medium, initial pH, atmosphere composition, incubation temperature and time, and prebiotics on growth and production of antimicrobial substances. Growth and antimicrobial substances production of L. brevis BK11 were significantly higher in MRS broth than in BHI or M17 broth. The production of cell mass, lactic acid, and bacteriocin by BK11 strain was at maximum in MRS broth adjusted to pH 6.0. Aerobic and microaerobic conditions were favored cell growth and antimicrobial substances production than anaerobic condition. Biomass and lactic acid production and antimicrobial substances activity of BK 11 were significantly better at 30 and $37^{\circ}C$ than at $25^{\circ}C$. Growth of the strain BK11 entered the stationary growth stage at 24 h after inoculation, and decreased after 36 h. Antimicrobial activities of cell-free culture supernatant and bacteriocin solution were highest when cultured in MRS broth with an initial pH 6.0 for 24-30 h at $37^{\circ}C$. In addition, the highest cell number and lactic acid and bacteriocin production were recorded in the presence of 1 and 2% (w/v) fructooligosaccharide (FOS), however, inulin and raffinose did not affect biological and physicochemical characteristics and antimicrobial activities of L. brevis BK11 cultures. According to these results, production of antimicrobial substances by L. brevis KB11 was closely associated with cell density. Under optimal conditions for antimicrobial substances production, L. brevis BK11 effectively inhibited the growth of Helicobacter pylori ATCC 43504.

Variation of Chemical Components and Their Interaction with Isoflavones in Maturing Soybean Seeds

  • Kim Sun-Lim;Lee Young-Ho;Yun Hong-Tae;Moon Jung-Kyung;Park Keum-Yong;Chung Jong-Il
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.50 no.4
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    • pp.291-300
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    • 2005
  • This study was focuses on the variation of isoflavone contents during seed development and their interaction with major chemical components such as protein, amino acids, saccaharides, lipid and fatty acids. During maturing, lipid, protein, and amino acid contents in soybean seeds showed the highest values at R7 stages, but isoflavone contents were increased until R8 stage. It was noted that malonyl glucosides $(64.2\%)$ are predominant forms among conjugated isoflavones followed by glucosides $(30.7\%)$, acetyl glucosides $(4.1\%)$ and aglycones $(0.9\%)$. Sucrose and stachyose were presented as a major saccharide in soybean seeds. As maturing days progressed, they were constantly increased and the highest contents were observed at R8 stage. While small quantities of raffinose, fructose, glucose, maltose, DP3 (DP: degree of polymerization), DP6, and DP7 were detected. These results showed that saccharide composition at the beginning of seed development is primarily monosaccharides with little sucrose and oligosaccharides, but as maturing days proceeds, sucrose and starch increase with concomitant decrease in monosaccharides. Sucrose and stachyose were positively correlated with isoflavone (r=0.780, 0.764 at p<0.01, respectively), while fructose, glucose, maltose, and DP7 were negatively correlated (r=-0.651, -0.653, -0.602, and -0.586 at p<0.05, respectively). Soybeans at R8 stage were high in protein and amino acid, but low in free amino acid contents. Protein and amino acid contents showed positively significant correlations with isoflavone (r=0.571 and 0.599 at p<0.05, respectively), but free amino acid content were negatively correlation with isoflavone (r=-0.673, p<0.01). The lipid content reaches its final content relatively early stage of seed development and remains constant as compared with other chemical components. Among the fatty acids, although varietal difference was presented, stearic acid and linolenic acid were gradually decreased, while oleic and linoleic acid were increased as seed maturing progressed. Lipid was significantly correlated (r=0.754, p<0.01) with isoflavones. However, neither saturated fatty acid nor unsaturated fatty acids significantly affected the isoflavone contents of maturing soybean seeds.

Studies on the Quality Evaluation and Metal Content of Sanitary Canned Kamju Beverage (위생캔으로 제조한 감주 음료의 품질 평가와 금속물질 함량에 관한 연구)

  • 허윤행
    • Journal of environmental and Sanitary engineering
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    • v.15 no.2
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    • pp.18-26
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    • 2000
  • In order to improve the quality of Kamju(sikhae), Korean traditional sweat fermentated rice drink, Kamju made with laboratory fermenting from both covered barley malt and that added to commercial $\alpha-amylase$ and commercial sikhae can were investigated and analyzed ad follows 1. The amylase activities in L-malt(laboraory made malt), C-malt(commercial malt) were 25,065 units, 20,498 units at $60^{\circ}C$ and when heating at $60^{\circ}C$ for one hour their heat stability was getting lower and while at $70^{\circ}C$ after 2 hours both their heat stability was under 21%. 2. The external appearance, of all samples, in beating test, flipper, springer and leaker test were excellent and good valued. On the open test of sample can, head space was 7.9~9.0mm, net weight 247~250g, 432~435g, pH 4.96-5.76, Brix 13.2~14.0 and vacuum degree was 29~35cmHg. 3. The sucrose content of sample 1~5 was 8,57~10.01% highest ammount than sample 6(0.91%) 7(0.43%) and maltose content of sample 1~5 was 0.65~1.41% lower than 6 was 7.91% and 7 was 8,36%. The good traditional flavor texture and saccharides components content of glucose of sample 1~5 was 1.09~1.51, 6(3.86%), 7(3.97%), fructose content of sample 1~5 was 0.45~0.95%, 6(2.76%), 7(2.81%), maltotriose and raffinose of sample 1~5 were 0.04~0.291, 6(0.61%), 7(2.81%), and solid content of sample 1~5(3.1~3.8) was lower than 6(10.25%), 7(10.09%). 4. The result of sensory test as measured as lemone yellow of sample 6 and milky of sample 7, and weetness, flavor, color, traditional texture and sensory value for sample 1~5(3.1~3.8) was lower degree than that for 6(4.5), 7(4.0). 5. Among the heavy metals analyzed from sample cans, tin, iron and lead content for sample 1~5 were 31.95~36.71ppm, 5.84~6.39ppm and 0.075~0.09ppm while their content of sample 6.7, tin, 32.91ppm, 32.87ppm, iron 5.51ppm, 5.41ppm and lead, 0.074ppm, 0.079ppm.

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최소가공 과일의 갈변저해를 위한 저해제의 탐색

  • 박연주;최소영;김영언;문광덕
    • Proceedings of the Korean Society of Postharvest Science and Technology of Agricultural Products Conference
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    • 2003.04a
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    • pp.119.1-119
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    • 2003
  • 단체 급식의 증가와 더불어 편의식품을 선호하는 소비 경향에 맞추어 최소가공한 과일의 갈변저해를 위한 저해제를 탐색하였다. 실험에 사용한 과일은 배와 감으로 각각 박피하여 속심을 제거한 후 배는 8조각으로 절단하고 감은 1~2cm 두께로 slice 하였다. 저해제는 1% ascorbic acid, citric acid, NaCl, MgCl$_2$, CaCl$_2$, oxalic acid, EDTA, raffinose, salicylic acit, sporix를 사용하였다. 또한 열수 추출한 오약, 계피, 황기, 당기, 건칡, 진피, 천궁, 감초 등도 저해제로 사용하였다. 최소가공한 과일은 저해제에 3분간 침지한 후 물기를 제거하고 tray에 포장하여 실온에서 저장하면서 colorimeter를 이용하여 갈변도의 변화를 측정하였다. 갈변 정도를 나타내는 $\Delta$E 는 시간의 경과에 따라 점차 감소하는 경향을 보였으나 배의 경우 변화 정도가 크지 않았다. NaCl, MgCl$_2$, CaCl$_2$를 처리한 배의 갈변 정도가 가장 적었다. cysteine을 처리한 구에서는 황화합물 냄새가 났다. 또한 oxalic acid를 처리한 구에서는 연화가 빠르게 일어났으며 저장 시간이 경과함에 따라 수분이 용출되었다. 한약재는 자체 색으로 인해 배의 색변화를 증가시켰으며 이취가 발생했다. 또한 저장 시간이 경과함에 따라 건조가 빠르게 일어나 관능적으로도 좋지 않은 결과를 나타냈다. 감에서는 CaCl$_2$, MgCl$_2$를 처리한 구에서 색의 변화가 가장 적었으며 cysteine은 색변화는 적었으나 배의 경우과 같이 이취가 발생하였다. 한약제는 계피와 진피를 처리한 감에서 색의 변화를 저해하는 효과를 보였다.를 처리하였을 경우 당귀, 천문동을 처리한 구에서 효과를 보였다. oxalic acid는 갈변저해에는 효과적이었으나 시간이 경과함에 따라 수분의 용출이 증가하였으며 물성의 변화도 컸다. 또한 ZnCl$_2$를 처리한 구에서는 이취 발생과 수분 용출이 컸으며 색의 변화도 컸다.내었다. 고온 단시간 처리구의 경우 저온 장시간 처리구와는 달리 저장 1일 후부터 그 농도가 다소 상승되었다가 다시 감소는 경향을 보였다. 한편 열처리하여 냉각시킨 직후 및 저장1주 후 사과의 pH, 산도, 당도, 경도, 과육의 갈변도 등을 조사하였던 바 대조구 및 처리구간에 일부 항목에 있어서는 약간의 차이를 나타내었다. 대치할 수 있을 것이라고 사료된다. 수분함량은 기계적 검사보다 관능검사와 더욱 높은 상관관계를 나타냈다.내었다. 항균활성이 우수한 생약재를 농도별로 활성을 조사한 결과, 물 추출물과 10% Ethanol 추출물 모두 낮은 농도에서도 우수한 항균활성을 나타내었다.취와 함께 점질성 갈변물질이 생성되었다. 이와 같은 결과로 볼 때, BAAG의 처리는 BAAC의 경우보다 가격은 저렴하면서도 항균력은 우수한 천연 항균복합제재로써 농산물 식품원료에 적용하여 선도유지 기간을 연장할 수 있는 효과를 기대할 수 있었다. 과일 등의 포장제로서 이용할 가능성을 확인하였다.로 [-wh] 겹의문사는 복수 의미를 지닐 수 없 다. 그러면 단수 의미는 어떻게 생성되는가\ulcorner 본 논문에서는 표면적 형태에도 불구하고 [-wh]의미의 겹의문사는 병렬적 관계의 합성어가 아니라 내부구조를 지니지 않은 단순한 단어(minimal $X^{0}$ elements)로 가정한다. 즉, [+wh] 의미의 겹의문사는 동일한 구성요 소를 지닌 병렬적 합성어([$[W1]_{XO-}$ $[W1]_{XO}$ ]

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Antimicrobial Susceptibility and Biochemical Characteristics of Streptococcus suis Isolated from Diseased Pigs in Gyeongbuk Province (경북지방 환돈에서 분리한 Streptococcus suis의 생화학적 성상 및 약제감수성)

  • Choi, Seong-Kyoon;Kim, Seong-Guk;Kim, Young-Hoan;Choi, Jeong-Hye;Jo, Min-Hee;Cho, Gil-Jae
    • Journal of Life Science
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    • v.20 no.10
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    • pp.1549-1555
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    • 2010
  • Streptococcus suis is a worldwide pathogen of a variety of porcine infection and has also been described as a pathogen for humans. We studied biochemical characteristics, antimicrobial susceptibility, and identification of polymerase chain reaction (PCR) of S. suis isolated from diseased pigs in Gyeongbuk province from 2004 to 2009. Sixty-one isolates were identified as S. suis by biochemical characteristics and PCR from 40 farms. The biochemical characteristics of S. suis isolates were production of VP-negative, hippurate, esculin, and arginine decarboxylase-positive, and fermentation of carbohydrate was variable lactose, trehalose, inulin, and raffinose, which was typeable 11 phenotype. In an antimicrobial susceptibility test, the majority of isolates were highly susceptible to amoxicillin/clavulanic acid, ampicillin, cephalothin, cefoperazone and florfenicol, while being highly resistant to streptomycin, kanamycin, amikacin, neomycin, erythromycin, clindamycin, and tetracycline. The isolates were divided into 11 phenotypes of biochemistry. By using PCR, the 16S-rRNA gene DNA fragment was detected at 304 bp from all of isolates. These results may provide the basic information needed to establish strategies for the prevention of S. suis infection in pigs.

Molecular Cloning and Expression of a Novel Protease-resistant GH-36 $\alpha$-Galactosidase from Rhizopus sp. F78 ACCC 30795

  • Yanan, Cao;Wang, Yaru;Luo, Huiying;Shi, Pengjun;Meng, Kun;Zhou, Zhigang;Zhang, Zhifang;Yao, Bin
    • 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.