Lactic acid bacteria (LAB) producing phenyllactic acid (PLA), which is known as antimicrobial compound, was isolated from button mushroom bed and the isolated LAB was identified to Lactobacillus casei by 16 rRNA gene sequence analysis. Cell-free supernatant (CFS) from L. casei was assessed for both the capability to produce the antimicrobial compound PLA and the antifungal activity against three fungal pathogens (Rhizoctonia solani, Botrytis cinerea, and Collectotricum aculatum). PLA concentration was investigated to be 3.23 mM in CFS when L. casei was grown in MRS broth containing 5 mM phenylpyruvic acid as precursor for 16 h. Antifungal activity demonstrated that all fungal pathogens were sensitive to 5% CFS (v/v) of L. casei with average growth inhibitions ranging from 34.58% to 65.15% (p < 0.005), in which R. solani was the most sensitive to 65.15% and followed by C. aculatum, and B. cinerea. The minimum inhibitory concentration (MIC) for commercial PLA was also investigated to show the same trend in the range of 0.35 mg mL-1 (2.11 mM) to 0.7 mg mL-1 (4.21 mM) at pH 4.0. The inhibition ability of CFS against the pathogens were not affected by the heating or protease treatment. However, pH modification in CFS to 6.5 resulted in an extreme reduction in their antifungal activity. These results may indicate that antifungal activities in CFS was caused by acidic compounds like PLA or organic acids rather than protein or peptide molecules.

Halitosis is caused by consumption of certain foods or drinks and production of volatile sulfur compounds (VSCs) by periodontopathogens. VSCs-related halitosis is not easily removed using mechanical or chemical therapies such as dental floss, plaque control and mouth rinse. Lactobacillus are known to be probiotics and stimulate immune systems of human. Furthermore, L. casei ATCC 334 and L. rhamnosus GG have an effect on protection of dental caries in vitro studies. The aim of this study was to investigate effect of Lactobacillus on halitosis by Fusobacterium nucleatum- and Porphyromonas gingivalis- producing VSCs and to analyze inhibitory mechanism. The periodontopathogens were cultivated in the presence or the absence Lactobacillus, and the level of VSCs was measured by gas chromatograph. For analysis of inhibitory mechanisms, the susceptibility assay of the spent culture medium of Lactobacillus against F. nucleatum and P. gingivalis was investigated. Also, the spent culture medium of Lactobacillus and periodontopathogens were mixed, and the emission of VSCs from the spent culture medium was measured by gas chromatograph. L. casei and L. rhamnosus significantly reduced production of VSCs. L. casei and L. rhamnosus exhibited strong antibacterial activity against F. nucleatum and P. gingivalis. The spent culture medium of L. casei inhibited to emit gaseous hydrogen sulfide, methyl mercaptan and dimethyl sulfide from the spent culture medium of periodontopathogens. However, the spent medium of L. rhamnosus repressed only dimethyl sulfide. L. casei ATCC 334 may improve halitosis by growth inhibition of periodontopathogens and reduction of VSCs emission.

Canine parvovirus (CPV2) is one of the most virulent virus causing acute hemorrhagic enteritis and myocarditis in dogs. Infection mainly caused by the ingestion of virus through the mucosal route. Therefore, induction of mucosal immunity is essential in prevention of Canine Parvovirus (CPV2) infection. For safe and effective delivery of viral antigens to the mucosal immune system, a novel surface antigen display system for lactic acid bacteria using the poly-γ-glutamic acid synthetase A protein (pgsA) of Bacillus subtilis as an anchoring matrix was applied in order to display CPV2 antigen on the surface of the recombinant L. casei. Recombinant fusion proteins comprised of pgsA and the capsid protein (VP2-S1) showed stable expression in Lactobacillus casei. Surface localization of the fusion protein was verified by cellular fractionation analyses. Oral and nasal inoculations of recombinant L. casei into mice resulted in high levels of serum immunoglobulin G (IgG) and mucosal IgA, as demonstrated by ELISA using recombinant VP2-S1 proteins. Mice receiving intranasal immunization mounted higher antibody response than those receiving oral immunization. These results indicate that mucosal immunization with recombinant L. casei expressing CPV2 VP2-S1 protein on its surface provides an effective means for elicitation of strong antibody responses against CPV 2 VP2-S1.

This research was carried out to find herbal preservatives for Makgeolli, as Makgeolli loses its commercial value due to overproduced acidic materials. When Makgeolli was kept at 25℃ to find the changes in acidity, total microbial cell number, yeast cell number, and bacterial species variety, a sudden increase of acidity as well as the disappearance of yeast cells occurred at day 6, and Makgeolli was changed to complete off-flavor. Acetobacter pasteurianus is the main acidifier in Makgeolli and shows a synergy effect in acid formation when cultured in combination with Lactobacillus casei. Among 12 herbs, the ethanol extract of Sutellaria baicalensis showed antimicrobial activity against A. pasteurianus, whereas the ethanol extract of Coptidis rhizoma showed antimicrobial activity against L. casei. Makgeolli added with Sutellaria baicalensis extracts demonstrated a lower acidity than that with Coptidis rhizoma extracts, which indicates that the inhibition of an acetic acid former is more important than that of a lactic acid former in Makgeolli preservation. Sutellaria baicalensis extracts prolonged the shelf life of Makgeolli by 1～2 weeks at a minimal inhibitory concentration (0.63 ㎎/㎖) during storage at 10℃.

This research was carried out in order to discover acid-forming bacteria during fermentation of Makgeolli, as Makgeolli loses its commercial value due to overproduced acidic materials. In Makgeolli kept at 25℃, a sudden increase of acidity as well as the disappearance of yeast cells occurred at day 6, whereas the total cell count and bacterial type remained unchanged; the result implies that a succession of bacterial types, including acid forming bacteria, occurred. Two acidforming bacteria were isolated from acidified Makgeolli and were identified as Acetobacter pasteurianus and Lactobacillus casei. When fresh and heat-treated Makgeolli were inoculated with Acetobacter pasteurianus and/or Lactobacillus casei, the greatest amount of acid was formed in Makgeolli inoculated with Acetobacter pasteurianus and Lactobacillus casei and also in Makgeolli with Acetobacter pasteurianus alone. This result indicates that Acetobacter pasteurianus is the main acidifier; furthermore, it shows the synergy effect in acid formation with Lactobacillus casei.

발효식품이 유해곰팡이에 의한 발암물질(aflatoxin)생성에 미치는 억제효과에 관한 연구의 일환으로 유산균 및 유산균 발효유가 Aspergillus parasiticus ATCC 15517의 성장과 aflatoxin 생성에 미치는 영향을 실험하였다. 발효유를 일정 농도별로 첨가한 YES 배지에서 Asperigillus parasiticus를 배양하여 그 성장과 배양물의 변화를 관찰하고 HPLC에 의하여 aflatoxin을 분석하였다. 그 결과 배양말기에 대조군에 비하여 건조 균체량, 배양물의 PH, 그리고 aflatoxin 생성량 등이 낮게 나타났다(p<0.05). Aflatoxin B₁은 48.6~58.1%가 감소되었으며 G₁은 29.8~34.2%가 감소되었다. 이 발효유의 발효에 사용된 유산간균(Lactobacillus casei)과 A. parositicus를 변형 APT 배지에서 혼합배양한 결과 A. parasitious 단독배양의 경우에 비하여 균체량이 배양 5일째까지는 현저하게 억제되었으나 배양 말기에는 유의한 차이를 보이지 않았다. 또한 배양 말기에 단독배양의 경우보다 pH가 훨씬 감소되고(p<0.05) aflatoxin의 생성량도 감소되었다. 이로부터 발효유는 유해곰팡이인 A. parasiticus의 성장과 aflatoxin 생성을 억제시키는 효과를 가짐을 알 수 있으며, 이는 발효에 관여한 미생물의 경쟁뿐만 아니라 유산균의 대사산물에 의한 영향으로 보여진다.