Korean Fermented Food Research Articles

Microbial β-Galactosidase of Pediococcus pentosaceus ID-7: Isolation, Cloning, and Molecular Characterization.

Pediococcus pentosaceus ID-7 was isolated from kimchi, a Korean fermented food, and it showed high activity for lactose hydrolysis. The β-galactosidase of P. pentosaceus ID-7 belongs to the GH2 group, which is composed of two distinct proteins. The heterodimeric LacLM type of β-galactosidase found in P. pentosaceus ID-7 consists of two genes partially overlapped, lacL and lacM encoding LacL (72.2 kDa) and LacM (35.4 kDa). In this study, Escherichia coli MM294 was used for the production of LacL, LacM, and LacLM. These three types of recombinant proteins were expressed, purified, and characterized. The specific activities of LacLM and LacL were 339 and 31 U/mg, respectively. However, activity was not detected with LacM alone. The optimal pH of LacLM and LacL was pH 7.5 and pH 7.0, and the optimal temperature of LacLM and LacL was 40°C and 50°C, respectively. The optimal temperature changes indicate that LacLM is able to achieve higher activity at a relatively lower temperature. LacLM was strongly activated by Mg2+, Mn2+, and Zn2+, which was not true for LacL. Consistent with this, EDTA strongly inactivated LacLM and LacL, but the presence of reducing agents did not dramatically alter the activity. Taken together, multiple alignment of amino acid sequences and phylogenetic analysis results of LacL and LacM of P. pentosaceus ID-7 suggest the evolution of LacL into LacLM and that the use of divalent metal ions results in higher activity.

A proteomic approach for rapid identification of Weissella species isolated from Korean fermented foods on MALDI-TOF MS supplemented with an in-house database

Weissella are obligate heterofermentative lactic acid bacteria belonging to the Leuconostocaceae family. Some Weissella can be found in salted and fermented foods, such as kimchi and jeotgal, and plays an important role in the fermentation process. In the present study, for the first time, a rapid and accurate identification method for Weissella species from kimchi and jeotgal was developed based on MALDI-TOF MS, supplemented with an in-house database. Of the 135 Weissella spectra aligned with the MALDI bioTyper database, 56 isolates (41.5%) yielded no reliable identification results with low log scores (<1.7). After registering the spectra of six Weissella reference strains, all of the isolates were correctly identified, of which 113 (83.7%) and 22 (16.3%) were identified at the species and genus level, respectively. Moreover, a dendrogram generated by protein profiles of the different Weissella species clearly presented distinctive clusters, and PCA analysis separated the spectra of Weissella species into four clusters. In comparing food origins, different Weissella species were identified from two fermented foods. W. soli and W. cibaria were isolated from kimchi, while W. thailandensis and W. halotolerans were isolated from jeotgal. The results of our proteomic approach confirm that the MALDI bioTyper database, with our in-house Weissella database, is sufficient for Weissella identification. The MALDI-TOF MS method provides fast and reliable discrimination between different species in the genus Weissella and, therefore, will be useful for safety control in fish farms or in the production of fermented foods. This method can also be applied to the control of opportunistic pathogenic Weissella in human clinical infections.

Isolation and Characterization of a Bacteriophage Preying an Antifungal Bacterium.

Several Bacillus species were isolated from rice field soils, and 16S rRNA gene sequence analysis showed that Bacillus cereus was the most abundant. A strain named BC1 showed antifungal activity against Rhizoctonia solani. Bacteriophages infecting strain BC1 were isolated from the same soil sample. The isolated phage PK16 had an icosahedral head of 100 ± 5 nm and tail of 200 ± 5 nm, indicating that it belonged to the family Myoviridae. Analysis of the complete linear dsDNA genome revealed a 158,127-bp genome with G + C content of 39.9% comprising 235 open reading frames as well as 19 tRNA genes (including 1 pseudogene). Blastp analysis showed that the proteins encoded by the PK16 genome had the closest hits to proteins of seven different bacteriophages. A neighbor-joining phylogenetic tree based on the major capsid protein showed a robust clustering of phage PK16 with phage JBP901 and BCP8-2 isolated from Korean fermented food.

Complete Genome Sequence of Pediococcus pentosaceus Strain wikim 20, Isolated from Korean Kimchi

Pediococcus pentosaceus strain wikim 20 is a lactic acid bacterium that was isolated from kimchi, a representative traditional Korean fermented food. Here, we announce the complete genome sequence of P. pentosaceus strain wikim 20 consisting of a 1,830,629-bp chromosome and provide a description of its annotation.

Properties of a Fibrinolytic Enzyme Secreted by Bacillus amyloliquefaciens RSB34, Isolated from Doenjang.

Nine bacilli with fibrinolytic activities were isolated from doenjang, a traditional Korean fermented soy food. Among them, RSB34 showed the strongest activity and was identified as Bacillus amyloliquefaciens by 16S rRNA and recA gene sequencing. During growth on LB up to 96 h, RSB34 showed the highest fibrinolytic activity (83.23 mU/μl) at 48 h. Three bands of 23, 27, and 42 kDa in size were observed when the culture supernatant was analyzed by SDS-PAGE and 27 and 42 kDa bands by fibrin zymography. The gene encoding the 27 kDa fibrinolytic enzyme AprE34 was cloned by PCR. BLAST analyses confirmed that the gene was a homolog to genes encoding AprE-type proteases. aprE34 was overexpressed in Escherichia coli BL21(DE3) using pET26b(+). Recombinant AprE34 was purified and examined for its properties. The Km and Vmax values of recombinant AprE34 were 0.131 ± 0.026 mM and 16.551 ± 0.316 μM/l/min, respectively, when measured using an artificial substrate, N-succinyl-ala-ala-pro-phe-p-nitroanilide. aprE34 was overexpressed in B. subtilis WB600 using pHY300PLK. B. subtilis transformants harboring pHYRSB34 (pHY300PLK with aprE34) showed higher fibrinolytic activity than B. amyloliquefaciens RSB34.

Assessment of Bacillus subtilis SN7 as a starter culture for Cheonggukjang, a Korean traditional fermented soybean food, and its capability to control Bacillus cereus in Cheonggukjang

The aim of this study was to develop a bacteriocin-producing QPS (Qualified Presumption of Safety) status Bacillus strain as a starter culture for fermented soybean foods as well as food preservation with improved safety and organoleptic qualities. Strain SN7 isolated from Meju, a fermented soybean product of Korea, was selected based on antimicrobial activity against pathogens such as B. cereus and production of bacteriocin. The selected strain SN7 was identified as B. subtilis. Safety assessments of B. subtilis SN7 (hemolytic activity, virulence traits, and antibiotic resistance) indicated that B. subtilis SN7 does not present a risk to human health, and this new isolate can be confidently recommended as a safe starter culture for the food industry. Cheonggukjang, a traditional fermented soybean food of Korea, fermented using B. subtilis SN7 showed high amino-type nitrogen content with good sensory characteristics. As a controlling agent of B. cereus in culture medium and in a food model of Cheonggukjang, B. subtilis SN7 efficiently inhibited growth of B. cereus vegetative cells and inactivated germination of B. cereus spores. Furthermore, B. cereus was not detected in Cheonggukjang stored at 4 °C for 6 months. B. cereus was easily detected in fermented food during storage at low temperature, as endo-spores of B. cereus are resistant to various treatments such as heat treatment and can grow even at 4–5 °C. These results suggest that bacteriocin-producing B. subtilis SN7 has great potential as a controlling agent of B. cereus and as a starter culture in Cheonggukjang.

Antibiotic susceptibilities and characteristics of Bacillus licheniformis isolates from traditional Korean fermented soybean foods

The antibiotic susceptibilities and characteristics of 94 Bacillus licheniformis strains isolated from traditional Korean fermented soybean foods were assessed. The minimum inhibitory concentration tests revealed that all strains were susceptible to gentamicin, kanamycin, tetracycline, and vancomycin, and that antibiotic resistances were expressed in a strain-specific manner. The resistances of B. licheniformis to chloramphenicol and streptomycin were established as intrinsic characteristics. The advent of erythromycin-resistant strains was attributed to the acquisition of a plasmid-mediated erythromycin resistance determinant. All strains sustained growth on tryptic soy agar (TSA) containing 14% NaCl (w/v), but 51.1% of strains did not grow at 15% NaCl. All strains exhibited protease and lipase activities on TSA but both activities decreased as the NaCl concentration increased. The function of both enzymes in high-salt soybean fermentation is therefore uncertain.

Gracilibacillus kimchii sp. nov., a halophilic bacterium isolated from kimchi.

A novel halophilic bacterium, strain K7(T), was isolated from kimchi, a traditional Korean fermented food. The strain is Gram-positive, motile, and produces terminal endospores. The isolate is facultative aerobic and grows at salinities of 0.0-25.0% (w/v) NaCl (optimum 10-15% NaCl), pH 5.5-8.5 (optimum pH 7.0-7.5), and 15-42°C (optimum 37°C). The predominant isoprenoid quinone in the strain is menaquinone-7 and the peptidoglycan of the strain is meso-diaminopimelic acid. The major fatty acids of the strain are anteisio-C15:0, iso-C15:0, and, C16:0 (other components were < 10.0%), while the major polar lipids are diphosphatidylglycerol, phosphatidylglycerol, phosphatidylcholine, and three unidentified lipids. A phylogenetic analysis of 16S rRNA gene sequence similarity showed that the isolated strain was a cluster of the genus Gracilibacillus. High levels of gene sequence similarity were observed between strain K7(T) and Gracilibacillus orientalis XH-63(T) (96.5%), and between the present strain and Gracilibacillus xinjiangensis (96.5%). The DNA G+C content of this strain is 37.7 mol%. Based on these findings, strain K7(T) is proposed as a novel species: Gracilibacillus kimchii sp. nov. The type strain is K7(T) (KACC 18669(T); JCM 31344(T)).

Safety and technological characterization of coagulase-negative staphylococci isolates from traditional Korean fermented soybean foods for starter development

To select starters for the production of meju and doenjang, traditional Korean fermented soybean foods, we assessed the safety and technological properties of their predominant isolates, Staphylococcus saprophyticus, Staphylococcus succinus and Staphylococcus xylosus. Phenotypic antibiotic resistance, hemolysis and biofilm formation were strain-specific. None of the S. succinus isolates exhibited antibiotic resistance or hemolytic activities. Thirty-three selected strains, identified through safety assessments of 81 coagulase-negative staphylococci (CNS) isolates, produced cadaverine, putrescine, and tyramine, but not histamine in the laboratory setting. The production of these three biogenic amines may, however, be insignificant considering the high levels of tyramine produced by the control, Enterococcus faecalis. The 33 CNS strains could grow on tryptic soy agar containing 21% NaCl (w/v), exhibited acid producing activity at 15% NaCl, and expressed strain-specific protease and lipase activities. S. succinus 14BME1, the selected starter candidate, produced significant amounts of benzeneacetic acid, 2,3-butanediol, trimethylpyrazine, and tetramethylpyrazine through soybean fermentation.

Isolation and Structural Characterization of an Oligosaccharide Produced by Bacillus subtilis in a Maltose-Containing Medium

Among 116 bacterial strains isolated from Korean fermented foods, one strain (SS-76) was selected for producing new oligosaccharides in a basal medium containing maltose as the sole source of carbon. Upon morphological characterization using scanning electron microscopy, the cells of strain SS-76 appeared rod-shaped; subsequent 16S rRNA gene sequence analysis revealed that strain SS-76 was phylogenetically close to Bacillus subtilis. The main oligosaccharide fraction B extracted from the culture supernatant of B. subtilis SS-76 was purified by high performance liquid chromatography. Subsequent structural analysis revealed that this oligosaccharide consisted only of glucose, and methylation analysis indicated similar proportions of glucopyranosides in the 6-linkage, 4-linkage, and non-reducing terminal positions. Matrix-assisted laser-induced/ionization time-of-flight/mass spectrometry and electrospray ionization-based liquid chromatography-mass spectrometry/mass spectrometry analyses suggested that this oligosaccharide consisted of a trisaccharide unit with 1,6- and 1,4-glycosidic linkages. The anomeric signals in the 1H-nuclear magnetic resonance spectrum corresponded to α-anomeric configurations, and the trisaccharide was finally identified as panose (α-D-glucopyranosyl-1,6-α-D-glucopyranosyl-1,4-D-glucose). These results suggest that B. subtilis SS-76 converts maltose into panose; strain SS-76 may thus find industrial application in the production of panose.

Korean traditional Chungkookjang improves body composition, lipid profiles and atherogenic indices in overweight/obese subjects: a double-blind, randomized, crossover, placebo-controlled clinical trial.

Chungkookjang is a Korean representative fermented soybean food. In this study, we investigated the effect of Korean traditional Chungkookjang compared with placebo on body composition, dyslipidemia and risk factors for atherosclerosis in overweight/obese subjects. This double-blind, randomized, controlled crossover trial was conducted on 120 overweight/obese subjects, aged 19-29 years. Subjects were randomly divided into a Chungkookjang (n=60) or a placebo (n=60) group. After 12 weeks, the groups were crossed over for an additional 12 weeks. During the intervention period, subjects were asked to maintain their usual diet and activity and not to take any functional foods or dietary supplements. The anthropometric measures, lipid profiles and atherogenic indices were determined at baseline and at the end of each 12-week period. The anthropometry measurements, percentage body fat, lean body mass, waist circumference and waist-to-hip ratio of women in the Chungkookjang group were significantly improved compared with the placebo group. Lipid profiles and high-sensitivity C-reactive protein of women in Chungkookjang were significantly improved. The atherogenic indices of apolipoprotein B/apolipoprotein A1 decreased in both the placebo and the Chungkookjang group, and it also decreased below 0.55 for all the men and women in the Chungkookjang group. In conclusion, these results suggest that supplementation with Chungkookjang may improve body composition and risk factors for cardiovascular disease in overweight and obese adults.

Characterization and Antibacterial Mode of Action of Lactic Acid BacteriumLeuconostoc mesenteroidesHJ69 from Kimchi

This study characterized a lactic acid bacterium Leuconostoc mesenteroides HJ69 at biochemical and molecular level, isolate from traditional Korean fermented food Kimchi. Cell free supernatant (CFS) of L. mesenteroides HJ69 exhibited significant (P < 0.05) antibacterial effect as diameters of inhibition zones (14.34 ± 0.31 – 18.21 ± 0.23 mm) against the tested foodborne pathogenic bacteria with minimum inhibitory concentration and minimum bactericidal concentration values found in the range of 250–2,000 and 500–2,000 μg/mL, respectively. Further, CFS of L. mesenteroides HJ69 revealed its mode of action on membrane integrity as confirmed by the increased release of potassium ions (800 and 750 mM/L), loss of 260-nm absorbing materials (3.85 and 3.77 OD), and increase in relative electrical conductivity (8.6 and 8.2%) against two selected bacteria L. monocytogenes KCTC-3569 (Gram-positive) and Escherichia coli O157:H7 (Gram-negative), respectively. The above findings hypothesize that L. mesenteroides HJ69 compromised its mode of action on membrane integrity, suggesting its enormous potential in the food industry. Practical Applications Asian countries have enormous use of fermented food products enriched with large populations of health-beneficial lactic acid bacteria (LAB). This study describes the isolation of a LAB strain Leuconostoc mesenteroides HJ69 from traditional Korean fermented food Kimchi along with its biochemical and molecular characterization. The cell free supernatant of L. mesenteroides HJ69 displayed remarkable antibacterial effect against various foodborne pathogenic bacteria with a hypothesized mode of action in several assays. These findings suggest that L. mesenteroides HJ69 could be an effective candidate for its practical application in food industry to control the proliferation of foodborne pathogenic bacteria as a natural antimicrobial agent.

Korean traditional fermented fish products: jeotgal

Jeotgal (醢) is a traditional Korean fermented food with thousands years of history with kimchi and other jang (fermented soybean products, 醬). The history was proved by research from historical literature and antique architecture. Jeotgal was developed along with jang (豆醬), fish jang (魚醬), meat jang (肉醬) as a part of jang (醬) up to the Chosun Dynasty and it was always offered during the ancestral rites or ceremonies. According to antique documents written by women, jeotgal had been used as seasonings or condiments that were popular especially for women rather than as food served for ancestral rites. In Southeast Asia and other countries, jeotgal uses varieties of fish and seafoods to provide rich and varied flavors, and thanks to the next generation sequencing technology, we can identify microorganisms that are involved in the fermentation process. Major microorganisms in jeotgal are Bacillus, Brevibacterium, Micrococcus, Pediococcus, Pseudomonas, Lactobacillus, Leuconostoc, and Halobacterium. Recently, much research on various health function of jeotgal has been conducted, reflecting increasing interest in the safety and the functionality of jeotgal. Many reports on functionalities of jeotgal such as supplying essential amino acids, and having antioxidant and antitumorgenic have been published recently. Because of the diverse flavor, types, and their function, jeotgal is expected to continue to develop as an important seasoning in the world sauce market.

항균력을 지닌 Bacillus 균주들을 종균으로 사용한 청국장에서 Bacillus cereus 억제

유해균인 Bacillus cereus와 독소 생성 곰팡이 증식 억제능력을 지닌 Bacillus amyloliquefaciens EMD17(EMD17)과 Bacillus subtilis EMD4(EMD4)를 단독 혹은 1:1로 대두에 접종하여 청국장을 제조하였다. 대조구로는 나토균주로 알려진 Bacillus subtilis KACC16450 균을 접종한 청국장을 제조하였다. B. amyloliquefaciens EMD17 접종 청국장에서는 오염시킨 B. cereus 증식이 즉시 억제되어 37℃ 발효 12시간에 검출되지 않았다. EMD4 접종 청국장에서도 B. cereus 증식 억제가 관찰되었으나 억제 정도는 약했다. 48시간 발효 후 청국장 시료들은 4℃에서 10일간 저장하였고 B. cereus는 EMD4 접종 청국장에서 검출되지 않았다. 그러나 대조구에서는 상당수 B. cereus 균수가 유지되었다. 청국장의 pH 값은 시료 간 유의차가 없었고 혈전용해능은 발효 중에는 EMD17을 접종한 청국장이 그리고 저장 중에는 나토균주를 접종한 것이 높았다. 총 페놀성 화합물도 나토균주 청국장에서 높았다. 이상 결과들에서 B. amyloliquefaciens EMD17 같은 균주들을 종균으로 사용함으로써 청국장에서 B. cereus 억제가 가능함을 알 수 있다.

한국전통발효식품 - 유익미생물의 잠재적인 자원과 응용

This review describes the diversity of Korean fermented foods and their significance as potential sources of probiotic bacteria. Fermented foods consumed in Korea are categorized according to their base material. Fermented foods such as kimchi, meju, doenjang, kangjang, jeotgal, and makgeolli are reported to have significant medicinal properties. These fermented products, which are consumed regularly by local people, are rich sources of beneficial microbes represented by several genera, including Weissella spp., Lactobacillus spp., Leuconostoc spp., Mucor, Penicillium, Scopulariopsis, Aspergillus, Rhodotorula, Candida, Saccharomyces, and Bacillus, as well as lactic acid bacteria. Fermented foods are now taken beyond the boundaries of their use as mere side dishes and are used significantly as a functional as well as medicinal foods. Fermented foods are a rich source of potential natural substances with antioxidant, anticancer, anticholesteric, antiobesitic, and antiaging properties, so that traditional fermented foods used as food supplements can impart health benefits. Publication of scientific studies on the dietary benefits of various fermented foods and growing consciousness about the potential health benefits of traditional fermented food are reflected in the scores of reports currently available in this field. Food microbiologists now have abundant opportunities to explore Korean traditional fermented foods for the isolation of new bacterial strains and to evaluate the potential applications of these strains through microbiological research.

Lentibacillus kimchii sp. nov., an extremely halophilic bacterium isolated from kimchi, a Korean fermented vegetable.

A Gram-positive, aerobic, non-motile and extremely halophilic bacterial strain, designated K9(T), was isolated from kimchi, a Korean fermented food. The strain was observed as endospore-forming rod-shaped cells showing oxidase and catalase activity. It was found to grow at 10.0-30.0% (w/v) NaCl (optimum, 15.0-20.0%), pH 7.0-8.0 (optimum, pH 7.5) and 15-40°C (optimum, 30°C). The polar lipids of strain K9(T) were identified as phosphatidylglycerol, three unidentified phospholipids and an unidentified glycolipid. The isoprenoid quinone was identified as menaquinone-7. The major cellular fatty acids (>20% of the total) were found to be anteisio-C15:0 and anteisio-C17:0. The cell wall peptidoglycan composition was determined to contain meso-diaminopimelic acid. The G+C content of genomic DNA was determined to be 48.2mol%. Phylogenetic analysis based on the 16S rRNA gene sequence revealed that the isolated strain is closely related to Lentibacillus salinarum AHS-1(T) (96.7% sequence similarity). Based on its phenotypic, chemotaxonomic and phylogenetic data, strain K9(T) is considered to represent a novel species of the genus Lentibacillus, for which the name Lentibacillus kimchii sp. nov., is proposed. The type strain is K9(T) (=KACC 18490(T)=JCM 30234(T)).

Microbial Community Structure of Korean Cabbage Kimchi and Ingredients with Denaturing Gradient Gel Electrophoresis.

Kimchi is a traditional Korean fermented vegetable food, the production of which involves brining of Korean cabbage, blending with various other ingredients (red pepper powder, garlic, ginger, salt-pickled seafood, etc.), and fermentation. Recently, kimchi has also become popular in the Western world because of its unique taste and beneficial properties such as antioxidant and antimutagenic activities, which are derived from the various raw materials and secondary metabolites of the fermentative microorganisms used during production. Despite these useful activities, analysis of the microbial community present in kimchi has received relatively little attention. The objective of this study was to evaluate the bacterial community structure from the raw materials, additives, and final kimchi product using the culture-independent method. Specifically, polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) was used to analyze the 16S rRNA partial sequences of the microflora. One primer set for bacteria, 341F(GC)-518R, reliably produced amplicons from kimchi and its raw materials, and these bands were clearly separated on a 35-65% denaturing gradient gel. Overall, 117 16S rRNA fragments were identified by PCR-DGGE analysis. Pediococcus pentosaceus, Leuconostoc citreum, Leuconostoc gelidum, and Leuconostoc mesenteroides were the dominant bacteria in kimchi. The other strains identified were Tetragenococcus, Pseudomonas, Weissella, and uncultured bacterium. Comprehensive analysis of these microorganisms could provide a more detailed understanding of the biologically active components of kimchi and help improve its quality. PCR-DGGE analysis can be successfully applied to a fermented food to detect unculturable or other species.

Inhibition of Bacillus cereus Growth and Toxin Production by Bacillus amyloliquefaciens RD7-7 in Fermented Soybean Products.

Bacillus cereus is a gram-positive, rod-shaped, spore-forming bacterium that has been isolated from contaminated fermented soybean food products and from the environment. B. cereus produces diarrheal and emetic toxins and has caused many outbreaks of foodborne diseases. In this study, we investigated whether B. amyloliquefaciens RD7-7, isolated from rice doenjang (Korean fermented soybean paste), a traditional Korean fermented soybean food, shows antimicrobial activity against B. cereus and regulates its toxin gene expression. B. amyloliquefaciens RD7-7 exhibited strong antibacterial activity against B. cereus and inhibited the expression of B. cereus toxin-related genes (groEL, nheA, nheC, and entFM). We also found that addition of water extracts of soybean and buckwheat soksungjang (Korean fermented soybean paste made in a short time) fermented with B. amyloliquefaciens RD7-7 significantly reduced the growth and toxin expression of B. cereus. These results indicate that B. amyloliquefaciens RD7-7 could be used to control B. cereus growth and toxin production in the fermented soybean food industry. Our findings also provide a basis for the development of candidate biological control agents against B. cereus to improve the safety of fermented soybean food products.

Genome Sequence of Bacillus glycinifermentans TH008, Isolated from Ohio Soil.

The genome sequence of an Ohio soil isolate, TH008, was determined. The sequence reveals a close relationship between TH008 and domesticated Bacillus glycinifermentans strains found in a traditional Korean fermented soybean food.

발효식품 추출물과 미생물을 활용한 참깨 종자전염성 병 방제

참깨 ì¢ ìžì „ì—¼ì„± 병해를 ë°©ì œí•˜ê¸° 위하여 다양한 ìœ ìš©ë¯¸ìƒë¬¼ì„ í•¨ìœ í•˜ëŠ” 김치, ê³ ì¶”ìž¥, 된장, 간장, 막걸리를 채집하여 식품 추출물을 ì¡°ì œí•˜ê³ ì¢ ìžì „ì—¼ì„±ë³‘ì›ê· ì— 대한 ì–µì œ 효과를 조사하였다. 추가로 ì‹í’ˆìœ ëž˜ 미생물의 참깨 ì¢ ìžì „ì—¼ì„± 병해 ì–µì œ 효과를 in vitro 및 in vivo 조건하에서 조사하였다. 50개 식품추출물에 처리한 참깨 ì¢ ìžë¥¼ ìƒ¤ë ˆìƒì— 처리하였을 때 20개 식품추출물이 ìœ ë¬˜ì©ìŒì¦ìƒì„ 92% 이상 ê°ì†Œì‹œì¼°ê³ , 7개 식품 추출물 처리는 온실조건에서 지상부 출현율을 58.3-66.8% 향상시키는 것으로 나타났다. 여러 발효식품으로부터 얻은 218 ì¢ ì˜ 발효미생물 중 7ì¢ ì˜ ì¢ ìžì „ì—¼ì„± ë³‘ì›ê· ì— 대하여 높은 í•­ê· í™œì„±ì„ 보였던 29ê· ì£¼ë¥¼ 처리하여 ì¢ ìžì†Œë íš¨ê³¼ë¥¼ ê²€ì • 한 ê²°ê³¼, 20ê· ì£¼ê°€ 병 ì–µì œ 효과가 있는 것으로 나타났다. 20ê· ì£¼ 중 13ê· ì£¼ëŠ” 모썩음 증상을 ìœ ì˜ì„± 있게 감소시켜 ê±´ì „ì£¼ìœ¨ì„ 향상시키는 것으로 나타났다. 16S rDNA region의 염기서열분석을 통해 in vitro 및 in vivo에서 í•­ê· í™œì„± 및 ì¢ ìžì†Œë íš¨ê³¼ë¥¼ 보인 ì‹í’ˆìœ ëž˜ ìœ ìš©ë¯¸ìƒë¬¼ 16ì¢ ì— 대한 ë™ì •ì„ 수행하였다. ê·¸ ê²°ê³¼ 14ê· ì£¼ëŠ” Bacillus 속 ì„¸ê· ìœ¼ë¡œ, 막걸리로부터 분리된 2ê· ì£¼ëŠ” Saccharomyces 속으로 ë™ì •ë˜ì—ˆë‹¤. 발효식품에서 분리된 ì„ ë°œ ê· ì£¼ëŠ” 대부분 B. amyloliquifaciens 인 것으로 나타났다. ì„ ë°œ ê· ì£¼ 중 B. amyloliquefaciens Gcj2-1와 B. amyloliquefaciens Gcj3-1를 ì½”íŒ ì œë¡œ bentonite, kaolin, talc 및 zeolite를 첨가 혼화하여 ì œì œí™”í•˜ì˜€ì„ 때 두 ê· ì£¼ 공히 talc 와 kaolin ì œí˜•ì´ ì–µì œ 효과가 ìš°ìˆ˜í•˜ì˜€ê³ ê·¸ 다음이 zeolite ì œí˜•ì´ì—ˆë‹¤. 그러나 bentonite ì œí˜•ì˜ 경우에는 타 ì œí˜•ì— 비해 ì–µì œ 효과가 낮게 나타났다. ì„ ë°œëœ ë°œíš¨ì‹í’ˆìœ ëž˜ 미생물은 in vitro와 온실조건에서 참깨 ì¢ ìžì „ì—¼ì„±ë³‘ ë°©ì œì— 있어서 효과 ì ì¸ 것으로 확인되었다. 이상의 결과를 근거로 국내산 발효식품 추출물 및 발효식품 ìœ ëž˜ 미생물은 참깨 ì¢ ìžì†Œë ìš© ë°©ì œì œë¡œì„œ ì‚¬ìš©ë ìˆ˜ 있을 것으로 생각된다.