Edible biopolymer films were developed from the exopolysaccharides (EPS) extracted from Weissella confusa 113-2. The optimum composition for film formation was determined using the response surface analysis with the explanatory variables of the EPS (0.5-5.5%) and glycerol (0.5-5.5%) concentrations and the response variable of film elastic modulus (EM). The mass ratio of distilled water to solids was set constant (14:1). Tensile strength (TS), percentage elongation at break (%E), EM, water vapor permeability (WVP) of EPS films were evaluated. The glass transition temperatures of the films were also determined by a dynamic mechanical analysis. The optimum mass ratio of EPS to glycerol was 0.754:0.375. The WVP, TS, %E, and EM of the film under the optimal composition were 3.53±0.21 g·mm/kPa·h·m2, 7.03±0.49 MPa, 84.82±12.31%, and 62.03±6.93 MPa, respectively. The glass transition temperature varied from 54 to 83 °C. The EPS film has the potential to be applied to food products as an edible film with physical and barrier properties comparable to other biopolymer edible films.

Acetic acid bacteria strains were isolated from a variety of fermented foods and fallen fruits. Among them, the strain MAK88, whose acetic acid fermentation ability, acid-tolerance, and alcohol-tolerance were high, was selected and identified as Acetobacter orientalis. A seed culture of A. orientalis MAK88 was inoculated into onion juice, and the optimum conditions of acetic acid fermentation was investigated. The optimum initial concentration of ethanol in onion juice was 5% (v/v) and in that condition, acidity was 4.31% at 144 h of fermentation. The optimum initial concentration of acetic acid was 1% and the final acidity was 5.32%. The optimum fermentation temperature was determined to be 28oC. The most appropriate preparation method of onion juice was to heat the onion at 121oC for 15 min and produce juice with pressure followed by filtering, and then sterilization at 121oC for 15 min. Prepared onion juice was used for fermentation without dilution.

Glucansucrase is an enzyme classified as a glycoside hydrolase (GH) 70 family, which catalyzes the synthesis of glucooligosaccharides with a low molecular weight using sucrose as a donor of D-glucopyranose and maltose as a carbohydrate acceptor. In this study, glucansucrase-producing lactic acid bacteria strain was isolated from the fermented foods collected in traditional markets, and the optimum conditions for the oligosaccharide production were investigated. The strain CCK940 isolated from Chinese cabbage kimchi was selected as an oligosaccharide-producing strain due to its high glucansucrase activity, with 918.2 mU/mL, and identified as Leuconostoc lactis. The optimum conditions for the production of oligosaccharides using Leu. lactis CCK940 were to adjust the initial pH to 6.0, add 5% (w/v) sucrose and 10% (w/v) maltose as a donor and acceptor molecules, respectively, and feed 5% (w/v) sucrose at 4 and 8 h of cultivation. When Leu. lactis CCK940 was cultured for 12 h at optimum conditions, at least four oligosaccharides with a polymerization degree of 2-4 were produced.

Glucansucrase is an enzyme classified as a glycoside hydrolase (GH) 70 family, which catalyzes the synthesis of various glucans and glucooligosaccharides with a low molecular weight using sucrose as a donor of D-glucopyranose and maltose as a carbohydrate acceptor. Oligosaccharides are indigestiable carbohydrate with low calories, which have various positive effects on the health of intestinal track. In this study, oligosaccharide produced from Leuconostoc lactis CCK940 was purified and its prebiotics effect was investigated. Leuconostoc lactis CCK940 which isolated from Chinese cabbage kimchi produced oligosaccharide using 5%(w/v) sucrose as a donor and 3%(w/v) maltose as an acceptor. Oligosaccharide produced was purified using Bio-gel P2 column chromatography and DPs of the resulting oligosaccharide were ranged from 3 to 7. When the prebiotics effects of freeze dried oligosaccharide were examined, 1%(w/v) of purified oligosaccharide added to carbohydrate-free MRS showed good prebiotics effects on Lactobacillus plantarum KCCM 12116, L. reuteri KCTC40417, and Bifidobacterium animalis KCCM1209.

For preliminary molecular typing, PCR-based fingerprinting using random amplified polymorphic DNA (RAPD) is the method of choice. In this study, 14 bacterial strains were isolated from different Korean food sources, identified using 16S rRNA gene sequencing, and characterized through RAPD-PCR. Two PCR primers (239 and KAY3) generated a total of 130 RAPD bands, 14 distinct PCR profiles, 10 polymorphic bands, one monomorphic band, and four unique bands. Dendrogram-based analysis with primer 239 showed that all 14 strains could be divided into seven clades out of which clade VII had the maximum of seven. In contrast, dendrogram analysis with the primer KAY3 divided the 14 L. citreum strains into four clades out of which clade IV consisted of a maximum of 10 strains out of 14. This research identified and characterized bacterial populations associated with different Korean foods. The proposed RAPD-PCR method, based on sequence amplification, could easily identify and discriminate the lactic acid bacteria species at the strain-specific level and could be used as a highly reliable genomic fingerprinting tool.

For preliminary molecular typing, PCR-based fingerprinting using random amplified polymorphic DNA (RAPD) is the method of choice. In this study, 14 bacterial strains were isolated from different Korean food sources, identified using 16S rRNA gene sequencing, and characterized through RAPD-PCR. Two PCR primers (239 and KAY3) generated a total of 130 RAPD bands, 14 distinct PCR profiles, 10 polymorphic bands, one monomorphic band, and four unique bands. Dendrogram-based analysis with primer 239 showed that all 14 strains could be divided into seven clades out of which clade VII had the maximum of seven. In contrast, dendrogram analysis with the primer KAY3 divided the 14 L. citreum strains into four clades out of which clade IV consisted of a maximum of 10 strains out of 14. This research identified and characterized bacterial populations associated with different Korean foods. The proposed RAPD-PCR method, based on sequence amplification, could easily identify and discriminate the lactic acid bacteria species at the strain-specific level and could be used as a highly reliable genomic fingerprinting tool.

Alginate lyase from Streptomyces violaceoruber was purified by DEAE sephacel chromatography and SP sepharose chromatography. The specific activity of the purified enzyme was 14.6 units/mg protein, representing a 40.6-fold purification of the crude extract. The final preparation thus obtained showed a single band on Tricine-SDS polyacrylamide gel electrophoresis whose molecular weight was determined to be 23.3 kDa. The polyMG block of sodium alginate was hydrolyzed by the purified alginate lyase and then separated by activated carbon column chromatography and bio gel P-2 gel filtration. The main hydrolysates were composed of hetero type M/G-oligosaccharides with the degrees of polymerization (D.P.) being 6 and 8. To investigate the effects of hetero type M/Goligosaccharides from the sodium alginate on the growth of some intestinal bacteria, cells were cultivated individually on the modified-MRS medium containing D.P. 6 and 8 M/G-oligosaccharides. B. longumgrew 4.25-fold and 6.44-fold more effectively by the treatment of D.P. 6 and 8 M/G-oligosaccharides compared with those of standard MRS medium. In addition, B. bifidumgrew 3.3-fold and 5.4-fold more effectively by the treatment of D.P. 6 and 8 M/G-oligosaccharides. In conclusion, D.P. 8 was more effective than D.P. 6 hetero M/G-oligosaccharides as regards the growth of Bifidobacteriumspp. and Lactobacillus spp. Key words: hetero M/G-oligosaccharides, Streptomyces violaceoruber

Alginate lyase from Streptomyces violaceoruber was purified by DEAE sephacel chromatography and SP sepharose chromatography. The specific activity of the purified enzyme was 14.6 units/mg protein, representing a 40.6-fold purification of the crude extract. The final preparation thus obtained showed a single band on Tricine-SDS polyacrylamide gel electrophoresis whose molecular weight was determined to be 23.3 kDa. The polyMG block of sodium alginate was hydrolyzed by the purified alginate lyase and then separated by activated carbon column chromatography and bio gel P-2 gel filtration. The main hydrolysates were composed of hetero type M/G-oligosaccharides with the degrees of polymerization (D.P.) being 6 and 8. To investigate the effects of hetero type M/Goligosaccharides from the sodium alginate on the growth of some intestinal bacteria, cells were cultivated individually on the modified-MRS medium containing D.P. 6 and 8 M/G-oligosaccharides. B. longumgrew 4.25-fold and 6.44-fold more effectively by the treatment of D.P. 6 and 8 M/G-oligosaccharides compared with those of standard MRS medium. In addition, B. bifidumgrew 3.3-fold and 5.4-fold more effectively by the treatment of D.P. 6 and 8 M/G-oligosaccharides. In conclusion, D.P. 8 was more effective than D.P. 6 hetero M/G-oligosaccharides as regards the growth of Bifidobacteriumspp. and Lactobacillus spp

Fermented food consists of a variety of lactic acid bacteria (LAB) that are also used in the industry as starter cultures. This genus comprises of different species that find importance as a preservatives in food like meat and sausages. Likewise, Lactobacillus brevis has been recognized as GRAS and is a potential probiotic strain extensively being used on an industrial scale. Since such bacteria are directly related to human health, there has been a need to identify and characterize them on the molecular level. In this study, LAB was identified and characterized from various fermented food samples available in South Korea. Two types to PCR-based molecular typing methods were used to analyze the 13 Lactobacillus brevis isolates, of which one was based on difference in the banding patterns originated on agarose gel and the other was related to the sequence analyses of various housekeeping genes in the particular strain. The former rep-PCR technique used three primers namely, REP, ERIC and (GTG)5 that amplified repetitive sequences in the genome and provided characteristic fingerprint profile for each isolate. This clustered the strains in 3 groups with the help of UPGMA method of clustering distinguishing between closely related strains. However, the latter multilocus sequencing typing (MLST) technique provided definite identification of the strains. A set of 7 housekeeping genes were determined as groEL, gyrB, rpoA, rpoB, pheS, recA and dnaK. These genes were amplified and sequenced and were subjected to comparative analysis. Discrete allelic profiles and 13 sequence types (STs) were resolved and minimum spanning tree (MST) was constructed, revealing the genetic relatedness among the isolates. On comparing the results from both the techniques, MLST proved to generate accurate and precise fingerprints owing to the sequence analysis of conserved genes thus providing a scope for research in the monitoring related species.

Glucansucrases are members of GH70 family of glycoside hydrolases and mainly produced by lactic acid bacteria such as Leuconostoc, Lactobacillus, and Weisella species. Glucansucrases use sucrose as a substrate to synthesize glucans as well as to produce oligosaccharides by transferring glucose moiety from sucrose onto acceptor carbohydrate molecules. It is known that some oligosaccharides produced by the reaction of glucansucrase show functional properties such as prebiotics and immunostimulatory activities. In this study, lactic acid bacteria which produce glucansucrase were screened from a variety of fermented foods, vegetables, and fruits collected from various area of Korea. Among 149 lactic acid bacteria strains which were isolated using BCP agar plates, 8 strains were selected which showed high glucansucrase activities. By nucleotide sequence analysis of 16S rRNA gene, strain no. 26-4 and 109-4 were identified as Leuconostoc mesenteroides, strain no. 95-4 strain was L. lactis, strain no. 92-4 and 112-1 were Weissella kimchii, strain no. 94-1 and 95-1 strains were W. confusa, and strain no. 113 was W. cibaria. When the oligosaccharides produced by the reaction of glucansucrase were analyzed using Bio-LC after these lactic acid bacteria were cultured into sucrose-containing media, the chromatograms of all bacterial showed unidentified peaks which implied the newly synthesized oligosaccharides

Recently, many studies have shown that propolis has diverse health beneficial effects such as anti-cancer, anti-bacterial, anti-inflammation, and anti-oxidant activities. Water-insolubility of propolis has made it extremely difficult in manufacturing health beneficial beverages containing propolis. In this study, the solubility of propolis was dramatically improved by entrapping with beta-cyclodextrin and the prebiotics effect of water-soluble propolis on the yogurt fermentation was investigated. Lactobacillus pentosus SC60 was selected as a starter for the yogurt fermentation due to its high DPPH radical scavenging activity. Low-fat milk which was supplemented with different concentrations of propolis (0.01, 0.05, 0.1, or 0.5% (w/v)) was fermented with L. pentosus SC60 for 30 h and the viable cell number, pH, and titratable acidity were checked. The rates of decrease in pH and increase in titratable acidity of yogurt after 12 h of fermentation were the highest when 0.5% (w/v) propolis were supplemented. Meanwhile pH and titratable acidity of yogurt without propolis reached 4.54 and 0.73%, respectively, most slowly after 30 hoffer mentation when compared with other yogurts supple mented with propolis. The viable cell number of L. pentosus SC60 in yogurt with 0.5% (w/v) propolis increased with highest rate and it exceeded 9 log CFU/mL at 8 h of fermentation, but that of yogurt with no propolis was only 8.54 log CFU/mL at 30 h of fermentation. This results showed that L. pentosus SC60 grew very well in the presence of water-soluble propolis and it is proposed that propolis acts as prebiotics

The genus Pediococcus belongs to the lactic acid bacteria and includes 15 species which are used in the food industry as both starter and probiotic cultures. The importance of Pediococcus spp. is due to their use as starter cultures in fermented meat as well as to their presence as the natural microbiota in vegetables. The availability of P. pentosaceus in the food industry increases the need for reliable molecular techniques for strain identification. To date, the reliable molecular methods for definite identification at strain level of microorganisms used in food industry has not been developed. Molecular identification based on suitable marker genes could be a promising alternative to conventional molecular typing methods such as ribotyping. In this study, the applicability of seven housekeeping genes gyrB, pyc, pgm, leuS, glnA, and dalR in combination with the pgi gene in multilocus sequence typing of P. pentosaceus was assessed. Sequencing and comparative analysis of sequence data were performed on 6 strains isolated from various vegetables. In addition to 17 sequence types, two new sequence types were identified and these fortified sequence types and seven marker genes allowed for a clear differentiation of the strains analyzed, indicating their applicability in molecular typing.

An alkalophilic microorganism, strain DK1122 producing an alkaline protease was identified. DK1122 was isolated from soil collected in central Korea. The strain DK1122 was Gram-positive, 0.7×2-4 μm in size, and its colony was yellowish white, The strain DK1122 was found to be spore-forming, catalase positive, oxidase positive, caseinolytic, and reduce nitrate to nitrite. The protease was produced aerobically on Horikoshi I agar medium (pH 9.0) with 1% (w/v) skim milk at 40°C for 24 h. Through 16S rRNA gene partial sequencing, the strain DK1122 had the 99.7% sequence similarity to 16S rRNA gene sequence of Bacillus pseudofirmus. Based on the biochemical and physiological properties as well as phylogenetic analysis, the isolated strain was named as Bacillus sp. DK1122. It is expected that Bacillus sp. DK1122 may be a promising candidate for a proudcer of an alkaline protease applicable to the food and detergent industries.

Bacillus agaradhaerens 균주의 xylanase 최적 생산조건을 검토하고 효소를 정제하여 그 특성을 확인하였다. 본 균주 의 xylanase 최적 생산조건은 1% CMC, 2% polypeptone, 0.1% K2HPO4, 0.02% MgSO4, 1% Na2CO3, 1% NaCl, pH 9.0이었으며, 종배양액을 1% (w/w)농도로 접종하여 40oC 에서 24시간 동안 160 rpm의 속도로 진탕배양하였을 때 xylanase를 최대로 생산하였다.

DEAE Sephadex column chromatography에 의해 Bacillussp. 유래 β-mannanase의 정제를 수행하여 비활성 21.57units/mL 정제배율 95.33배를 나타내었다. SDS-PAGE에 의한 단일밴드를 확인하였고, 분자량은 38.9kDa으로 결정되었다.정제효소에 의해 Picea abies galactosyl glucomannan을 가수분해하여 activated carbon column chromatography에 의해 당가수분해물을 분리 회수하여 TLC, FACE 및Timell’s method에 의해 중합도 8, 10으로 결정되었으며,Penicillum purpurogenum유래 정제 β-mannanase와 α-galactosidase를 이용한 enzymatic sequential action에 의해 2가지 가수분해산물 모두 hetero type galactosylglucomannooligosaccharides로 확인되었다. B. longum, B.bifidum, B. infantis, B. animalis, B. breve, B. adolessentis,B. auglutum의 생육활성에 대한 중합도 8, 10의 영향을 검토하기 위하여 modified-MRS 배지상에 탄소원으로 중합도8, 10를 대체하여 생육활성을 비교한 결과 B. animalis에서는 중합도 8 galactosyl glucomannooligosaccharide를 탄소원으로 대체한 경우 표준 MRS 배지와 비교하여 19.5배, 중합도 10에서 18.7배의 가장 우수한 생육활성을 나타내었으며, B. bifidum에 대해서는 중합도 8의 경우 15.3배와 중합도10에서 14.3배 그리고 B. longum에서는 중합도 8 galactosylglucomannooligosaccharide를 탄소원으로 대체한 경우 표준MRS배지와 비교하여 중합도 8에서 15.2배, 중합도 10에서13.9배의 상대활성을 우선적으로 나타내었다.

팽화홍삼에 존재하는 major ginsenoside를 minorginsenoside로 생물전환하기 위하여 β-glucosidase 활성이높은 유산균주를 탐색하였다. 된장으로부터 분리한 YLB 8균주가 β-glucosidase 활성이 가장 높았으며 Leuconostocmesenteroides로 동정되었다. L. mesenteroides YLB 8을MRS 배지에서 배양하였을 경우 최대의 β-glucosidase 효소 활성을 나타내기 위한 최적 배양 조건은 glucose 첨가량 1%(w/v), 초기 pH 8.0, 배양온도 30oC, 배양시간 16시간이었다. L. mesenteroides YLB 8을 이용하여 팽화홍삼추출액을 생물전환하였을 경우 최적 온도는 30oC었으며,팽화홍삼 추출액의 농도는 12oBx이었다. 팽화홍삼 추출액내에 존재하는 major ginsenoside인 Rb1과 Rb2는 minorginsenoside인 Rg3로 전환되었으며, Re는 Rg1으로 전환되었으며, 이 중 일부는 Rh1과 Rh2로 전환되었다.

pH의 변화는 10%, 15%, 20% 염도 고추장에서는 숙성기간이 지나면서 약간의 pH의 감소 현상이 일어나기는 했지만, 수치에 큰 변화는 없었다. 그러나 염도가 낮은 3%와6% 고추장은 급격한 pH의 감소를 보여주었다. 호기성 세균수는 숙성기간 동안 큰 변화가 없었지만, pH의 감소 경향과 비슷하게 3%와 6% 염도 고추장에서 생균수가 더 많이 감소했다. 모든 염도에서 B. licheniformis와 B. subtilis가 큰 비중으로 차지하였다. 그러나 염도가 낮은 3%와 6% 염도 고추장에서는 이외에도 다양한 균주가 확인되었다. 호염성 세균수의 변화 추이는 호기성 세균의 생균수 변화 추이와 비슷했다. 호염성 세균도 B. subtilis가 모든 염도에서 전 숙성기간 동안 가장 큰 비중으로 확인되었다. 그러나 호염성 세균에서는 B. licheniformis 대신 Peanibacillus 속이 많은 부분에서 지배균주였다. 이외에도 3% 염도 고추장에서는 숙성 기간 동안 다양한 균주가 확인되었다. 효모는분리 유무가 숙성기간에 따라 다르게 나타났는데, 이는 pH의 급격한 저하에 의해 낮은 pH에도 생육이 가능한 균주만 숙성후기에 분리되는 것으로 판단된다. 20% 이상의 염도에서는 pH보다 고염에 의한 영향으로 효모의 증식이 어려운 것으로 보인다. 모든 염도에서 Candida 속과 S.cerevisiae가 가장 많이 분리되었으며, 이외에도 다양한 균종이 확인되었다. 검출된 휘발성 향기성분은 ester류가 26종으로 가장 많고, 그 다음 alcohol류가 12종, ketone류가 6종등 총 70종이 검출되었다. 이 중 alcohol류의 ethanol, ester류의 ethyl acetate와 3-methyl-1-butyl acetate, ketone류의 3-hydroxy-2-butanone, acid류의 acetic acid 등이 10-40% 정도의 면적비율로 검출되면서 주요한 성분으로 나타났다. 효모의 대사산물이면서 고추장의 고소한 향미를 내는ethanol은 10%와 15%에서 상대적으로 높은 비율로 검출되었고, 과실향에 기여하는 ethyl acetate는 3%, 6%, 10% 염도 고추장에서 상대적으로 높게 나타났으며, 반대로 발효유의 중요한 향기성분인 3-hydroxy-2-butanone은 염도가높아질수록 높은 비율로 검출되었다. 바나나의 향기에 중요한 3-methyl-1-butyl acetate와 초산발효에 중요한 기여를하는 acetic acid는 3%와 6% 염도 고추장에서 매우 높은수치로 검출되면서 고추장의 이취를 내는 것으로 보인다.결론적으로, 일반적인 고추장의 미생물 균총과 휘발성 향미가 달라지지 않게 제조하기 위한 고추장의 최저 염도는10%가 가장 적절할 것으로 사료된다.

연구에서는 전분질 원료를 달리한 18종류의 입국을 제조하고 당화력을 측정, 각각의 입국과 동일한 원료를 첨가하여 제조한 막걸리의 품질 특성과 관능검사 및 저장성 실험 결과를 분석하였다. 각각의 입국으로 제조한 막걸리의 에탄올 함량은 9-13%까지 고루 분포되었으며 제조한 막걸리 No. 5(현미) 군에서 13%로 가장 높게 측정되어 막걸리의 에탄올 생성에 있어 현미군이 유리할 것으로 사료된다. 막걸리의 총 당량은 막걸리 No. 6(찹쌀) 군이 11.3%, 환원당 또한 마찬가지로 4.8의 가장 높은 수치를 보여 막걸리의 당 생성에 있어 찹쌀군이 유리할 것으로 사료된다. pH는 막걸리 No. 2(밀+보리) 군이 3.6으로 가장 낮게 측정되었으며 총산은 6.0-9.1 사이로 고루 분포하였으며, 막걸리를 5oC에 보관 하였을 때 산도의 증가는 크지 않았으며 28oC에서 보관 하였을 때 3일 동안 총산이 급격하게 증가 하였지만 전분질 원료간의 특징적인 차이는 보이지 않았다.

Pediococcus pentosaceus KC-007가 생산하는 박테리오신은 MRS 배지에서 대수 증식기 말기에 최대로 생산되어 세포 외로 분비되었다. 박테리오신 활성은 proteinase K,trypsin, chymotrypsin, pepsin, subtilisin A 등의 단백질 가수분해효소에 의해 완전히 저해되었으나, catalase, lysozyme,lipase, ribonuclease A, α-amylase에 의해서는 저해되지 않고 안정하게 유지되었다. 박테리오신은 pH 2부터 8까지 범위에서 활성이 완전하게 유지되었으며, 100oC에서 60분간 또는 autoclave 처리에 의해서도 활성이 유지되었다. 박테리오신활성은 acetone, chloroform, ethanol, hexane, isopropanol,methanol 등의 유기용매나 SDS, Tween 20 등의 계면활성제의 작용에 영향을 받지 않았다. 본 연구에서 사용한 박테리오신은 Bacillus cereus, Escherichia coli O157-H7,Listeria monocytogenes과 같은 식중독 세균에 대한 높은항균활성을 지니고 있으므로 식품 보존제로서의 이용성이높을 것으로 판단된다.

Pulsed electronic field(PEF) 처리에 의한 우유 단백질과 물리화학적 특성의 변화를 확인하기 위하여 원유, 탈지유, HTST, LTLT, UHT 우유를 PEF 처리하였다. 시료 중의 단백질을 SDS-PAGE로 확인하였을 때, PEF 처리에 의한 우유 단백질의 변성은 관찰할 수 없었다. Differential scanning calorimetry(DSC)로 우유 단백질의 열변성 정점 온도(Td)를 분석한 결과, 탈지유를 65oC에서 PEF 처리하였을 때 Td가 87.66oC에서 97.18oC로 증가하여 PEF 처리가 우유 단백질의 변성에 영향을 미치는 것을 확인하였다. PEF 처리에 의한 alkaline phosphatase, protease, lactoperoxidase의 잔존효소활성을 측정한 결과, 원유와 탈지유에서 alkalinephosphatase는 PEF 처리에 의해 효소활성이 감소하였다. 또한 protease와 lactoperoxidase의 활성은 PEF 처리에 의해 영향을 받지 않았다. 65oC에서 PEF 처리한 원유는 처리하지 않은 원유보다 높은 갈색도를 나타내었으나, 기타 우유는 PEF에 의한 유의적인 차이가 없었다. 우유를 PEF 처리하였을 경우 산도의 변화는 관찰되지 않았고 pH의 경우에도 PEF 처리 여부에 따라 유의적인 차이는 있었으나 크게 변화하지는 않았다.