본 연구에서는 벼 부산물인 미강을 이용한 글루텐-프리 조미소재를 개발하기 위하여 Aspergillus oryzae를 이용한 미 강의 발효조건을 평가하였다. Aspergillus oryzae의 종류에 따라 미강을 발효하여 발효액의 일반성분을 분석한 결과 조 단백, 총질소(TN), 아미노태질소(AN) 항목에서 MC-01 균주가 높은 함량을 나타내었다. 균주별 발효액의 SDS-PAGE 와 유리 아미노산 분석 결과에서도 저분자성 물질로의 분해율이 MC-01 균주에서 가장 우수한 것으로 평가되었다. MC-01 균주를 이용하여 미강을 13일까지 발효를 진행하면서 3, 5, 7, 10, 13일 경과 후 각각의 미강 발효액의 품질을 평가하였다. 발효 10일 시료에서 조단백, 총질소(TN), 아미노태질소(AN)의 함량이 가장 높게 나타났고, 총 유리아미노 산 함량은 발효 10일까지 증가하다 이후부터는 유사한 수준을 나타내었다. 관능평가 결과 색, 향, 감칠맛, 종합 기호도 에서 발효 10일 시료의 기호도가 가장 높게 나타났다. 따라서 조미소재 개발을 위한 미강의 발효는 MC-01 균주를 이 용하여 10일 동안 발효시키는 것이 적합할 것으로 사료된다.

In this study, we compared the organoleptic and other qualities of fermented milk containing 10 or 15% purple carrot extract that had either been previously fermented with Aspergillus oryzae or not fermented. Fermentation characteristics, pH, chromaticity, viscosity, viable cell counts, and sensory evaluations were measured. The pH and acid values did not differ between purple carrot extract fermented with Aspergillus oryzae and non-fermented extract. Viable cell counts were significantly higher in 15% purple carrot extract fermented with Aspergillus oryzae compared to the control after fermentation. Regarding characteristic changes, purple carrot extract fermented with Aspergillus oryzae group showed a lower red value but higher yellow value compared with non-fermented purple carrot extract due to heat-sterilization. Both fermented and non-fermented extract groups showed significantly increased viscosity compared to control. In the sensory evaluation, 15% purple carrot extract fermented with Aspergillus oryzae showed the highest score. In conclusion, addition of 15% purple carrot extract fermented with Aspergillus oryzae resulted in a superior fermented milk product.

It is possible to apply DNA sequencing data of A. oryzae RIB40 (Tominaga et al, 2006) to investigation of genomic structure of homologous gene cluster in 210 A. oryzae RIB strains. Using PCR technique, 210 A. oryzae RIB strains were easily classified into groups 1, 2, and 3, and others according to amplified patterns with seven aflatoxin homologous genes. Group 1 (122 strains, 58.1%) strains conserve intact aflatoxin biosynthesis gene homolog cluster. Group 2 (77 strains, 36.7%) and group 3 strains (9 strains, 4.3%) reveal large deletions of the aflatoxin gene homolog cluster, which is more than half. It is possible that the breakpoint within the cluster of group 2 strains would be near the ver-1gene, as described by Kusumoto et al. (2000). Two strains (0.9 %) that could not be classified into group 1, 2, and 3 were called "others". The majority of A. oryzae RIB strains (94.8 %) are categorized as groups 1 and 2. Murakami (1971) has evaluated 20 mycological characters of RIB strains, graded them from 1 to 6 and also proved no aflatoxin production in all strains. To examine the differences between group 1 and group 2 based on phenotype, analysis of variance was performed. Significant differences among 19 characters except for the aflatoxigenic character were recognized with 5 characters (length of stalk, color of old slant culture, roughness of conidia, coloration of hydroquinone, and pink color of conidia in medium with anisic acid). The length of stalk of group 1 was longer than that of group 2 at level of p<0.01 (data not shown). Therefore, this PCR analysis is a useful method for classification at intra-species level. Furthermore, it is safe for the food fermentation and enzyme industry to use A. oryzae especially groups 2 and 3 strains since these strains revealed absolute lack of aflatoxigenic ability at the molecular level. From the results of DNA sequencing analysis between A. oryzae RIB40 belonging to group 1 and RIB62 belonging to group 2, RIB62 shows a large deletion upstream of ver-1 homolog with more than half of the aflatoxin homologous gene cluster being missing. Adjacent to the deletion of the aflatoxin homologous gene cluster, RIB62 has a "unique sequence" of about 8-kb and a telomere. We investigated whether homologues of the unique sequence region of A. oryzae RIB62 were present in other group strains with Southern blot analysis. At first, we performed Southern blot analysis of 210 A. oryzae RIB strains with "no-hit" probe of unique sequence. The results showed that all group 2 strains had an identical size of signal of about 9.4-kb, while in other group strains different size of hybridizing signals from that of group 2 strains or with no signal were detected (data not shown).Subsequently, to confirm the presence of the unique sequence, Southern blot analysis with the four kinds of probe, which were derived from the unique sequence of RIB62 was performed for 16 selected strains from group 1, 2, and 3. Group 1 strains showed various signal patterns; double or single band(s) in most strains and with no signal in RIB40. In addition, the signal pattern of group 1 strains was different according to the probe used. However all group 2 strains showed an identical band of about 9.4 kbin all the cases when the four probes were used. In the group 3 strains, no signal was detected with the four probes. Therefore, 8-kb unique sequence of RIB62 is conserved in A. oryzae group 2 strains and present partially in some group 1 strains. To investigate the chromosomal position of the unique sequence, chromosomal Southern analysis was performed using four kinds of probe, US 1 to 4. Separation of the chromosomes of selected eight of A. oryzae group 1 and group 2 strains by clamped homogeneous electric field (CHEF) revealed different karyotype (data not shown). Among them, RIB62 showed a unique band of about 3 Mb, whereas other strains have no this chromosome. The detected signal(s) in A. oryzae group 1 strains were revealed in two or one chromosome(s) and in no signal in RIB40, while that of group 2 strains showed in single chromosome with four kinds of probes. The signal patterns of group 1 strains were different according to the probe used, while those of group 2 strains were identical. These results are almost identical to those of genomic Southern blot analysis To confirm the structure of the region flanking the partial aflatoxin homologous gene cluster in A. oryzae group 2 strains, we investigated the pattern of PCR amplification in 210 A. oryzae RIB strains with a set of primers designed to amplify between ver-1 and the unique sequence. The oligonucleotide primer for the ver-1 side was common to both RIB 40 and RIB62, while that of the unique sequence side was derived from RIB62. From the results of PCR with this set of primers, a fragment of about 1 kb was amplified from all group 2 strains and none of strains from other group generated PCR products. Therefore, it is possible to distinguish group 2 strains from other group strains with this set of primers. Southern blotting and PCR analysis resulted that all group 2 strains has the identical "unique sequence" and genomic structure of deletion including flaking region. In addition, this characterization of group 2 strains could be applied to distinguish this group strainsfrom the other group strains. The result of chromosomal Southern analysis (data not shown) suggested that the aflatoxin homologous gene cluster and the "unique sequence"existed on the same chromosome in groups 1 strains having these two portions. Taken together, A. oryzae group 2 strains might have differentiated from the ancestral strain due to chromosomal breakage. Although it is extremely difficult to determine the reason for the non-aflatoxigenicity of A. oryzae from the analysis of the genomic structure, this dissertation may provide basic molecular information for the profound approaches. In succession, further research on aflR protein activity or other related signal transduction pathway and the deleted aflatoxin biosynthesis gene homolog cluster of group 2 strains together with group 3 strains may help in clarifying the mechanism of the cluster deletion and differentiation.

본 연구는 Aspergillus Oryzae (AO) 및 Saccharomyces cerevisiae (SC)를 첨가하여 제조한 맥주박 위주 발효사료의 발효특성 및 영양학적 특성을 검토하기 위해 실시하였다. 시험구 처리는 시험사료에 AO 를 첨가하여 제조한 발효사료 처리구(FFAO), SC 를 첨가하여 제조한 발효사료 처리구(FFSC) 및 AO 와 SC 를 첨가하여 제조한 발효사료 처리구(FFAS)로 나누었다. 48시간 발효에 따른 조단백질 함량은 처리

본 연구는 Aspergillus oryzae(AO) 혹은 Saccharomyces cerevisiae(SC)를 첨가하여 제조한 맥주박 위주 발효사료의 급여가 한우의 반추위내 미생물에 미치는 영향을 조사하기 위해 실시하였다. 공시동물은 반추위 cannula가 장착된 한우 암소 2두를 이용하였다. 시험은 시판 배합사료 및 corn silage 급여구(대조구), 시판 배합사료 , AO 첨가 발효사료 및 com silage 급여구(TAO)와 시판 배합사료 ,

당화효소와 호정화효소의 생산능력이 높고 향도 좋은 Aspergillus oryzae L2와 밀가루로써 누룩을 제조할 때에 당화효소와 단백질분해효소의 생산을 위한 배양조건을 검토하였다. 밀가루를 가열처리 했을 때에는 날밀가루에 비하여 단백질분해효소의 생산은 증가되었으나 당화효소의 생산은 감소되었다. 밀가루에 0.5%의 염산을 함유하는 물을 급수했을 때에는 당화효소와 단백질분해호소의 생산이 현저하게 감소되었다. 급수비율은 밀가루에 대하여 28%일 때가 가장 좋았다. 국균을 접종한 후 즉시 성형을 하는 것보다 20시간 전 배양을 한 후에 성형을 하는 것이 당화효소의 생산에 좋았고 단백질분해효소의 생산은 성형을 하지 않은 것이 좋았다. 당화효소생산의 최적온도는 36℃이었고 단백질분해효소생산의 최적 온도는 28℃이었다.

In order to improve the quality of Takju, the effect of combined use of Aspergillus oryzae and Aspergillus kawachii in brewing was investigated. The quality of Takju which was brewed by the combined use of Aspergillus orytae CF7-koji and Aspergillus kawachii CF1-koji in equal amount was better as compared with that brewed by existing method using Aspergillus kawachii-koji only as koji. But the good result did not obtained when the koji was made by mixed culture of Aspergillus CF7 and Aspergillus kawachii CF1.

본 연구는 재래누룩에서 분리 동정한Aspergillus oryzae 83-10(AO)을 활용하여 원료조건에 따른 pellet형 누룩의 특성과 양조특성을 검토하였다. 실험 결과, 원료의 조건은 10분간 증자하여 살균․호화된 원료(S)가 pellet 누룩의 제조에 있어서 안정적이고, 높은 효소력의 생성에 더욱 적합한 것으로 나타났으며, 발효경과 32시간 때에 효소활성이 높고, 포자생성에 의한 포자 날림 등이 없어 작업성이 가장 우수하였다. 제조된 pellet 누룩 S-AO(32시간 배양)과 시판되고 있는 재래누룩 SH의 효소활성(dry base, unit/g)을 비교한 결과, glucoamylase 활성 S-AO(325.92) > SH(297.44), α-amylase 활성 S-AO(421.26) > SH(75.12) 및 acidic carboxypeptidase 활성 S-AO(3,803.8) > SH(278.6)으로 pellet 누룩이 재래누룩과 비교해 유의적으로 높은 수치를 나타내었으며, 누룩의 일반분석 결과에서는 SH 누룩이 pH, 총산, 아미노산도 모두 유의적으로 높은 수치를 나타내었다. 약주 양조특성 분석 결과 발효가 끝난 시점에 SH 술덧의 가용성고형물과 총산함량이 S-AO 술덧과 비교하여 유의하게 높게 나타났으며, S-AO 술덧은 아미노산 함량과 알코올 함량이 유의적으로 높게 나타났다. 알코올 함량 분석결과 S-AO 17.96%, SH 12.66%으로 큰 차이를 나타내어 발효 안정성과 수율에 있어서 S-AO 술덧이 높은 평가를 받았다. 발효경과 중 술덧의 효소활성은 누룩의 사용량을 쌀 사용량(g)×30 unit/g(glucoamylase)으로 효소활성 수치에 따라 달리 하였지만 그에 못 미치는 결과를 보였다. 술덧제조 직후 S-AO 26.69 unit/mL, SH 17.58 unit/mL으로 S-AO 시료는 비교적 근접한 수치를 나타내었지만 SH 시료는 상당량 부족한 모습을 보여 이러한 결과는 누룩의 품질 균일성에 의한 것으로 판단되어진다. 이러한 결과로 살균된 통밀을 pellet형 누룩으로 가공하여 국균을 배양하면 기존 재래식 누룩이나 입국에 비하여 품질을 떨어뜨리지 않으며, 제조효율도 상당히 높일 수 있을 것으로 기대할 수 있다.

The purpose of this study was to investigate the physiochemical properties of Doenjang was fermented by added with fungi and protease. The moisture content and pH of Doenjang added with protease (WP) were lower than those of control w/o protease while the contents of titratable acidity, reducing sugar, and amino-type nitrogen in WP were higher than control. The α-amylase activities of Doenjang added with single and mixed Protease B were the highest at 4 weeks of fermentation period and protease activity of WP was about 4 times higher than that of control. The 4-9 kinds of free amino acids (proline, isoleucine, leucine, and phenylalanine etc.) in WP was increased in comparison with control. The DPPH radical scavenging activity and total polyphenol content were higher in WP than control. Total aerobic bacterial and fungal numbers were decreased depending on fermentation time regardless of addition of protease. In conclusion, the protease can be used as additives improving the quality and taste of fermented Doenjang.

Koji (Aspergillus oryzae) is used to ferment crude cereals of wheat to make a traditional alcoholic drink called Makkolli and industrial materials. It’s quality varies depending on the wheat quality. Four domestic wheat varieties (Kosomil, Jokyungmil, Geumgangmil, Baegjungmil) were characterized. They were found similar in pH (6.02 to 6.08) and total acid (0.105 to 0.120%) contents. However, amino acid content of Gemgangmil was the highest (4.46%) and that of Baegjungmil was the lowest (3.72%). The total bacillus number was highest in Kosomil (333×103CFU/ml) and lowest in Gemgangmil (60×103CFU/ml). On the other hand, the fungus number was 47×105CFU/ml in Gemgangmil and the other varieties had similar quantity. The content of Alpha-amlyase was the highest (500.01unit/g) in Kosomil followed by Jokyungmil and Gemgangmil, and the lowest was in Baegjungmil (353.32unit/g). The content of Glucoamlyase was the highest in Geumgangmil (5105.0unit/g) followed by Jokyungmil and Kosomil, and the lowest was in Baegjungmil (3880.0unit/g). Acid protease was the highest in Kosomil (3515.15unit/g) followed by Geumgangmil and Baegjungmil, and the lowest in Jokyungmil (1280.5unit/g). From the result, Koji made from Kosomil was found to be of superior quality.

A. oryzae로 발효한 청미래덩굴(Smilax china L.)잎 발효차의 적정 발효기간을 확립하고자 비 발효(NF) 및 10, 20 및 30일간 발효(F10, F20, F30)시킨 차 1% 열수추출물(1 tea bag 기준)의 색상, 관능검사 및 total polyphenol(TP), total flavonoid(TF), 전자공여능(EDA), 철환원력(FRAP), 과산화물 생성 억제능(LPOIA)을 조사하였다. 또, 체내 활성 산소(ROS) 생성계 효소인 동시에 요통과 음주로 인한 간 손상유도 및 이로 인한 복부비만에 직간접적으로 관여하는 xanthine oxidase(XO) 및 aldehyde oxidae(AO)의 저해활성에 미치는 영향을 조사하였다. 색상과 spectrum(400～700nm)의 변화를 조사한 결과, NF는 연한 황색을 띠는 반면 F10∼F30에서는 엷은 적색을 띠었으며 F10의 색상이 가장 선명하였다. 향(aroma)과 밝기(brightness)에 대한 기호도는 비발효차와 발효차간의 유의차를 보이지 않았으나 맛 (taste)과 입에 닿는 감각(mouth feel) 및 종합적인 기호도 (overall acceptability)는 F10, F20 및 F30 간의 뚜렷한 차이를 보이지 않아 발효 10일이 이상적인 발효기간이라 사료된다. TP 함량은 NF에서 41.55 mg/g(dry basis)이었으나 발효에 따라 거의 비례적으로 감소하였으며 그 감소율은 발효 10일째 24.91%, 20일째 56.92%, 30일째 64.41%를 나타내었다. TF의 함량은 NF에서 27.33 mg을 나타내었으나 발효에 점차적으로 감소하여 F10 24.30 mg/g, F20 17.32 mg/g, F30 13.22 mg/g으로 감소하였다. 그러나 TP의 감소율이 TF의 경우에 비하여 커서 TF/TP 비율(%)은 발효에 따라 증가하는 경향을 나타내었다. EDA는 NF에서는 29.01%이었으나 F10에서는 NF에 비하여 17.14%가 감소하였으며, F20 및 F30에서는 각각 18.79% 및 23.20%가 감소하였다. FRAP(μ M Fe2+)는 NF 4.63, F10 4.30, F20 및 F30에서는 각각 3.77 및 3.47로 발효에 따라 감소하는 경향을 보였다. LPOIA는 NF에서는 39.86%이었으나 F10의 경우는 31.92%로 NF에 비하여 19.92%가 감소하였고 F20 및 F30는 NF에 비하여 각각 23.61% 및 28.38%가 감소하였다. NF 및 F10∼30의 1% 열수추출액이 생유 및 토끼 간 조직으로부터 부분정제한 XO와 AO의 활성에 미치는 영향을 조사한 결과, XO활성에는 비발효, 발효 모두에서 뚜렷한 영향을 미치지 않았다. 그러나 AO의 활성은 비발효, 발효 관계없이 38.09∼41.70% 범위로 억제하였으며, 이러한 억제는 경쟁적 저해현상에 기인되어 나타난 것으로 생각된다.