We compared the volatile profiles in soy sauce according to inoculation with Tetragenococcus halophilus and/or Zygosaccharomyces rouxii. Totals of 107 and 81 volatiles were respectively identified by using solidphase microextraction and solvent extraction. The various volatile compounds identified included acids, aldehydes, esters, ketones, furans and furan derivatives, and phenols. The major volatiles in the samples treated with T. halophilus were acetic acid, formic acid, benzaldehyde, methyl acetate, ethyl 2-hydroxypropanoate, 2-hydroxy-3-methyl-2-cyclopenten-1-one, and 4-hydroxy-3-methoxybenzaldehyde, while those in the samples inoculated with Z. rouxii were mainly ethanol, acetaldehyde, ethyl propanoate, 2/3-methylbutanol, 1-butanol, 2-phenylethanol, ethyl 2-methylpropanoate, and 4-hydroxy-2-ethyl-5-methyl-3(2H)-furanone. The results indicate that T. halophilus produced significant acid compounds and could affect the Z. rouxii activity, supporting the notion that yeasts and lactic acid bacteria respectively have different metabolic pathways of alcoholic fermentation and lactic acid fermentation, and produce different dominant volatile compounds in soy sauce.Key words: soy sauce; volatile compound; volatile profile; Zygosaccharomyces rouxii; Tetragenococcus halophilus Soy sauce, a soybean-based fermented food, is used in the cooking and seasoning of many Asian foods. Its popularity is increasing worldwide, mainly due to its characteristic flavor and umami taste.1) The industrial manufacturing process for soy sauce generally consists of three main steps: making koji, fermenting brine, and refining. Making koji involves inoculating cooked soybeans with Aspergillus oryzae (A. sojae) which produces such extracellular enzymes as proteases and amylases that hydrolyze proteins and polysaccharides in the soybeans. A brine mixture is then made by steeping the obtained koji in a salt solution. The brine mixture can be inoculated at this point with such a pure cultured lactic acid bacterium as Tetragenococus halophilus and such yeast as Zygosaccharomyces rouxii. Since these salt-tolerable lactic acid bacteria and yeasts are the predominant microorganisms during brine fermentation, adding pure cultured T. halophilus and Z. rouxii can accelerate and develop the formation of flavor in soy sauce.2,3) The high salt concentration also prevents the growth of undesirable microorganisms. The enzymes from the koji continuously break down the macromolecules in soybeans into smaller molecules such as sugars, peptides, and amino acids. During the growth of T. halophilus and Z. rouxii, diverse secondary metabolites, including the volatile flavor components, are formed via different metabolic pathways during lactic acid fermentation and alcoholic fermentation.3) The objective of brine fermentation is consequently to maximize both flavor production and hydrolysis of the macromolecules in soybeans.4) Nearly 300 volatile flavor compounds in soy sauce have been identified by many studies.5-7) The various volatiles identified in soy sauce...

The aim of this study was to identify volatile and agar-diffusible antifungal metabolites produced by Bacillus sp. G341 with strong antifungal activity against various phytopathogenic fungi. Strain G341 isolated from four-year-old roots of Korean ginseng with rot symptoms was identified as Bacillus velezensis based on 16S rDNA and gyrA sequences. Strain G341 inhibited mycelial growth of all phytopathogenic fungi tested. In vivo experiment results revealed that n-butanol extract of fermentation broth effectively controlled the development of rice sheath blight, tomato gray mold, tomato late blight, wheat leaf rust, barley powdery mildew, and red pepper anthracnose. Two antifungal compounds were isolated from strain G341 and identified as bacillomycin L and fengycin A by MS/MS analysis. Moreover, volatile compounds emitted from strain G341 were found to be able to inhibit mycelial growth of various phytopathogenic fungi. Based on volatile compound profiles of strain G341 obtained through headspace collection and analysis on GC-MS, dimethylsulfoxide, 1-butanol, and 3-hydroxy-2-butanone (acetoin) were identified. Taken together, these results suggest that B. valezensis G341 can be used as a biocontrol agent for various plant diseases caused by phytopathogenic fungi.

Restricted usage of chemical nematicides has led to development of environmentally safe alternatives. A culture filtrate of Aspergillus niger F22 was highly active against Meloidogyne incognita with marked mortality of second-stage juveniles (J2s) and inhibition of egg hatching. The nematicidal component was identified as oxalic acid by organic acid analysis and gas chromatography-mass spectroscopy (GC-MS). Exposure to 2 mmol/L oxalic acid resulted in 100% juvenile mortality at 1 day after treatment and suppressed egg hatching by 95.6% at 7 days after treatment. Oxalic acid showed similar nematicidal activity against M. hapla, but was not highly toxic to Bursaphelenchus xylophilus. The fungus was incubated on solid medium and dried culture was used for preparation of a wettable powder-type (WP) formulation as an active ingredient. Two WP formulations, F22-WP10 (ai 10%) and oxalic acid-WP8 (ai 8%), were prepared using F22 solid culture and oxalic acid. In a field naturally infested with M. incognita, application of a mixture of F22-WP10 + oxalic acid-WP8 at 1,000- and 500-fold dilutions significantly reduced gall formation on the roots of watermelon plants by 58.8 and 70.7%, respectively, compared to the non-treated control. The disease control efficacy of the mixture of F22-WP10 + oxalic acid-WP8 was significantly higher than that of a chemical nematicide, Sunchungtan (ai 30% fosthiazate). These results suggest that A. niger F22 can be used as a microbial nematicide for the control of root-knot nematode disease.

The antagonistic fungus Simplicillium lamellicola BCP has been developed as a microbial biopesticide that effectively controls the development of various plant diseases caused by both pathogenic bacteria and pathogenic fungi. Antibacterial bioassay-directed fractionation was used to isolate mannosyl lipids from S. lamellicola BCP, and the structures of these compounds were elucidated using spectral analysis and chemical degradation. Three novel mannosyl lipids were characterized and identified as halymecins F and G and (3R,5R)-3-O-β-D-mannosyl-3,5-dihydrodecanoic acid. Massoia lactone and (3R, 5R)-3-hydroxydecan-5-olide were also isolated from S. lamellicola BCP. The three novel compounds inhibited the growth of the majority of phytopathogenic bacteria that were tested, and halymecin F displayed the strongest antibacterial activity. Agrobacterium tumefaciens was the most sensitive to the three novel compounds, with IC₅₀ values ranging from 1.58 to 24.8 μg/mL. The ethyl acetate extract of the fermentation broth from the antagonistic fungus effectively reduced the bacterial wilt caused by Ralstonia solanacearum on tomato seedlings. These results indicate that S. lamellicola BCP suppresses the development of plant bacterial diseases through the production of antibacterial metabolites.

This is the first report on the suppression of PWD by OA. The result strongly indicates that PWD could be controlled by antibacterial antibiotic alone and a combination of antibacterial and nematicidal agents.