Traditional Fermented Food In Taiwan Research Articles

Isolation and characterization of lactic acid bacteria from xi-gua-mian (fermented watermelon), a traditional fermented food in Taiwan

Young watermelon fruit was peeled and pickled for fermentation to produce a unique fermented food named xi-gua-mian (fermented watermelon) in Taiwan. In this study, we investigated the LAB microflora in xi-gua-mian. A total of 176 LAB isolates were identified; 118 cultures were isolated from the xi-gua-mian sample collected from three different farmers markets and 58 from six young watermelon fruit samples. These isolates were characterized phenotypically and then divided into seven groups (A to G) by restriction fragment length polymorphism analysis, sequencing of 16S ribosomal DNA and other genotypic analysis. Lactobacillus plantarum was the most abundant LAB found in xi-gua-mian samples collected in southern Taiwan, Tainan City and Pediococcus pentosaceus was the most abundant LAB in northern Taiwan, Taoyuan County. We found that LAB stains are similar in samples collected in the same geographic region but significant variations were observed between samples collected among different regions. On the other hand, a greater LAB diversity was observed in the young watermelon fruit samples. In addition, 10 Lactococcus lactis subsp. lactis showed inhibitory activity against the indicator strain L. sakei subsp. sakei JCM 1157T. This is the first report describing the distribution and varieties of LAB existing in the xi-gua-mian and the young watermelon fruits.

Isolation, Identification and Characterization of Lactic Acid Bacteria From Shao-jiou-luo (Fermented Zoned Cerith), a Traditional Fermented Food in Taiwan

This study investigated the distribution of the native lactic acid bacteria (LAB) population in shao-jiou-luo (fermented zoned cerith), a popular traditional fermented food in Taiwan. A total of 129 cultures of LAB were isolated from shao-jiou-luo samples, and 14 cultures were isolated from its main raw material, zoned cerith. These isolates were divided into classes by phenotype and then into groups by restriction fragment length polymorphism analysis and sequencing of 16S ribosomal DNA. The identification results revealed 11 bacterial species belonging to five genera in shao-jiou-luo and three species belonging to two genera in zoned cerith. The most common bacterial genera in shao-jiou-luo were heterofermentative Weissella and Leuconostoc. On the other hand, Enterococcus aquimarinus was the most common species found in the raw zoned cerith sample. In conclusion, the results suggest that heterofermentative LAB species greatly contributed to the fermentation of shao-jiou-luo. This is the first report describing the distribution and varieties of LAB that exist in the shao-jiou-luo fermentation process and raw zoned cerith.

Lactococcus formosensis sp. nov., a lactic acid bacterium isolated from yan-tsai-shin (fermented broccoli stems)

A coccal-shaped organism, designated 516(T), was isolated from yan-tsai-shin (fermented broccoli stems), a traditional fermented food in Taiwan. 16S rRNA gene sequencing results showed that strain 516(T) had 98.9 % sequence similarity to that of the type strain Lactococcus garvieae NBRC 100934(T). Comparison of three housekeeping genes, rpoA, rpoB and pheS, revealed that strain 516(T) was well separated from Lactococcus garvieae NBRC 100934(T). DNA-DNA hybridization studies indicated that strain 516(T) had low DNA relatedness with Lactococcus garvieae NBRC 100934(T) (46.1 %). The DNA G+C content of strain 516(T) was 38.1 mol% and the major fatty acids were C16 : 0 (22.7 %), C19 : 0 cyclo ω8c (17.9 %) and summed feature 7 (29.0 %). Based on the evidence, strain 516(T) represents a novel species of the genus Lactococcus, for which the name Lactococcus formosensis sp. nov. is proposed. The type strain is 516(T) ( = NBRC 109475(T) = BCRC 80576(T)).

Isolation and characterization of lactic acid bacteria from Yan-tsai-shin (fermented broccoli stems), a traditional fermented food in Taiwan

To isolate, characterize and identify lactic acid bacteria (LAB) in yan-tsai-shin (fermented broccoli stems), a traditional fermented food in Taiwan. A total of 226 LAB were isolated; 111 cultures were isolated from samples collected from seven different markets and 115 from six fresh broccoli samples. These isolates were characterized phenotypically and then initially divided into nine groups (r1 to r9) using restriction fragment length polymorphism analysis and sequencing of 16S ribosomal DNA. Some isolates were further divided into four additional groups by other genetic analysis. The most common bacterial genera in yan-tsai-shin and fresh broccoli were Weissella, Lactococcus and Lactobacillus. Regional similarities in the LAB, with differences in diversity, were observed in this study. On the basis of phylogenetic analysis of 16S rRNA, rpoA, rpoB and pheS gene sequences, two strains were included in the genera Enterococcus and Lactococcus, respectively, and identified as potential novel species or subspecies. In addition, the novel enterococcal strain, and 33 L.lactis subsp. lactis and two Weissella cibaria strains were found to have bacteriocin-like inhibitory substance (BLIS) producing abilities. These results suggest that the LAB play important roles in the fermentation of yan-tsai-shin. This is the first report describing the distribution and varieties of LAB existing in yan-tsai-shin and fresh broccoli. In addition, two potential novel LAB species or subspecies and one potential novel BLIS were also found in this study.

Lactococcus taiwanensis sp. nov., a lactic acid bacterium isolated from fresh cummingcordia

One coccal strain, designated 0905C15(T), was isolated from fresh cummingcordia, which is the main ingredient of pobuzihi (fermented cummingcordia), a traditional fermented food in Taiwan. 16S rRNA gene sequencing results showed that strain 0905C15(T) had 98.22-98.82 % sequence similarity to that of the type strains of four Lactococcus lactis subspecies (L. lactis subsp. lactis BCRC 12312(T), L. lactis subsp. cremoris BCRC 12586(T), L. lactis subsp. hordniae BCRC 80474(T) and L. lactis subsp. tructae BCRC 80475(T)). Comparison of two housekeeping genes, recA and rpoB, revealed that strain 0905C15(T) was well separated from the reference strains of the genus Lactococcus. DNA-DNA hybridization studies indicated that strain 0905C15(T) had low DNA relatedness to the four Lactococcus lactis subspecies (9.7-15.24 %). The DNA G+C content of strain 0905C15(T) was 39.6 mol %. Based on the evidence, strain 0905C15(T) represents a novel species of the genus Lactococcus, for which the name Lactococcus taiwanensis sp. nov. is proposed. The type strain is 0905C15(T) ( = NBRC 109049(T) = BCRC 80460(T)).

Isolation and characterization of lactic acid bacteria from pobuzihi (fermented cummingcordia), a traditional fermented food in Taiwan

Lactobacillus pobuzihii is a novel species which has been previously found in pobuzihi (fermented cummingcordia), a traditional fermented food in Taiwan. However, the lactic acid bacteria (LAB) microflora in pobuzihi has not been studied in detail. In this study, LAB from pobuzihi were isolated, identified, and characterized. A total of 196 LAB were isolated; 79 cultures were isolated from the sample collected from a manufacturing factory, 38 from pobuzihi samples collected from 4 different markets, and 79 from 2 fresh cummingcordia samples. These isolates were characterized phenotypically and then divided into eight groups (A to H) by restriction fragment length polymorphism analysis and sequencing of 16S ribosomal DNA. Lactobacillus plantarum was the most abundant LAB found in most samples during the fermentation of pobuzihi. On the other hand, Enterococcus casseliflavus and Weissella cibaria were, respectively, the major species found in the two fresh cummingcordia samples. A potential novel species or subspecies of lactococcal strain was found. In addition, seven L. plantarum and five W. cibaria strains showed inhibitory activity against the indicator strain Lactobacillus sakei JCM 1157(T). This is the first report describing the distribution and varieties of LAB existing in the pobuzihi during its fermentation process and the final product on the market.

Isolation and characterisation of lactic acid bacteria from jiang‐gua (fermented cucumbers), a traditional fermented food in Taiwan

Jiang-gua (fermented cucumbers) is a popular traditional fermented food in Taiwan. The microflora of lactic acid bacteria (LAB) in jiang-gua have not been investigated in detail. In this study, LAB from jiang-gua were isolated, characterised and identified. A total of 103 LAB were isolated; 70 cultures were isolated from jiang-gua samples and 33 cultures were isolated from its raw substrate, cucumber. These isolates were mainly characterised phenotypically and then divided into seven groups (A-G) by restriction fragment length polymorphism analysis and sequencing of 16S ribosomal DNA. The isolates were identified as Enterococcus casseliflavus, Leuconostoc lactis, Leuconostoc mesenteroides, Lactobacillus pentosus, Lactobacillus plantarum, Lactobacillus paraplantarum, Lactococcus lactis subsp. lactis, Weissella cibaria and Weissella hellenica. The antibacterial activities of the isolates were determined and 11 Lc. lactis subsp. lactis strains showed inhibitory activity against the indicator strain Lactobacillus sakei JCM 1157(T) . Heterofermentative W. cibaria and Leu. lactis were the major LAB found in jiang-gua samples without soy sauce. In soy sauce-added samples, homofermentative L. pentosus and L. plantarum were the most abundant LAB. In addition, the results also suggested that HhaI and RsaI restriction enzymes could be applied to distinguish W. hellenica and Weissella paramesenteroides.

Diversity of lactic acid bacteria in sian‐sianzih (fermented clams), a traditional fermented food in Taiwan

Sian-sianzih (fermented clams) is a popular traditional fermented food in Taiwan. The lactic acid bacteria (LAB) microflora in sian-sianzih have not been studied in detail. In this study, LAB from sian-sianzih were isolated, characterized and identified. A total of 186 cultures of LAB were isolated from seven sian-sianzih samples and 29 cultures were isolated from its main raw substrate: clams. The identification results revealed up to 11 distinct bacterial species belonging to five genera in sian-sianzih, and three species belonging to two genera in clams. The most common bacterial genera in sian-sianzih were Lactobacillus and Weissella, followed by Leuconostoc, Pediococcus and Lactococcus. A regional similarity in LAB, with differences in diversity, was observed in the current study. On the other hand, Lactococcus lactis subsp. lactis was the most common species found in raw clam samples. The results also suggested that greater LAB diversity could be observed in wild clams than in cultured ones. Furthermore, antibacterial activities of the isolates were determined, and one Weisella hellenica strain showed inhibitory activity against the indicator strain Lactobacilluas sakei JCM 1157(T) . A sensory assessment of seven sian-sianzih samples was also performed and the results indicated that diversity of LAB has a great effect on its aroma and taste formation. The results demonstrate that various LAB species are distributed in sian-sianzih and have a great effect on the flavor of sian-sianzih.

Isolation and characterisation of lactic acid bacteria from yan-jiang (fermented ginger), a traditional fermented food in Taiwan.

Yan-jiang (fermented ginger) is a popular traditional fermented food in Taiwan. The lactic acid bacteria (LAB) microflora in yan-jiang has not been studied in detail. In this study, LAB from yan-jiang were isolated, characterised and identified. A total of 176 LAB were isolated; 160 cultures were isolated from yan-jiang samples and 16 cultures were isolated from raw ginger. These isolates were characterised phenotypically and then divided into nine groups (A to I) by restriction fragment length polymorphism analysis and sequencing of 16S ribosomal DNA. Lactobacillus sakei and Lactococcus lactis subsp. lactis were the major LAB found in the initial 2 days of fermentation without pickled plums; these species were mostly replaced by Weissella cibaria and L. plantarum after 3 days of fermentation. In the fermentation bucket with added pickled plums, W. cibaria was the most abundant LAB found during fermentation. The antibacterial activities of the isolates were determined. Twenty-four Lc. lactis subsp. lactis and 19 W. cibaria strains showed inhibitory activity against the indicator strain L. sakei JCM 1157(T) . Results demonstrate that various LAB species were more numerous when fermentation was carried out without pickled plums. LAB also had effects on the aroma and flavour of yan-jiang.

Isolation and characterization of lactic acid bacteria from jiang-sun (fermented bamboo shoots), a traditional fermented food in Taiwan

In this study, lactic acid bacteria (LAB) were isolated, characterized and identified from jiang-sun (fermented bamboo shoots; a traditional fermented food in Taiwan). Samples were collected at seven time intervals from a fixed fermenting bucket during the fermentation process of jiang-sun and its initial ingredients, dochi (fermented soybeans) and bamboo shoots. A total of 57 LAB cultures were isolated; 42 cultures were isolated from jiang-sun samples and 15 cultures were isolated from dochi and bamboo shoots. These isolates were characterized phenotypically and then divided into three groups (A-C) by restriction fragment length polymorphism analysis and sequencing of 16S ribosomal DNA. Alteration of microbial populations during the fermentation process was observed. While various LAB were found in the dochi and bamboo shoots, it was mostly replaced by Lactobacillus plantarum after 1 day of fermentation. Furthermore, the antibacterial activities of the isolates were determined, and one Enterococcus faecium strain showed inhibitory activity against all the indicator strains. Results suggest that L. plantarum is the main LAB present during the fermentation of jiang-sun. To our knowledge, this is the first report describing the distribution and varieties of LAB that exist in the jiang-sun fermentation process.

Isolation and characterization of lactic acid bacteria from Yan-dong-gua (fermented wax gourd), a traditional fermented food in Taiwan

Yan-dong-gua (fermented wax gourd; a traditional fermented food in Taiwan) samples were collected at five time intervals from a fixed fermenting bucket. Eighty-five cultures of lactic acid bacteria (LAB) were isolated from Yan-dong-gua samples, and the isolates were divided into classes by phenotype and then groups by restriction fragment length polymorphism analysis and sequencing of 16S ribosomal DNA. Phenotypic and biochemical characteristics identified two bacterial groups (A and B) and showed that the majority of the isolates were heterofermentative LAB. Weissella cibaria was the major LAB found during the initial fermentation of yan-dong-gua. However, it was mostly replaced by W. paramesenteroides after 5 days of fermentation. All isolated LAB were able to grow in MRS broth containing 6% NaCl. Furthermore, the antibacterial activities of the isolates were determined, and five of the isolates showed inhibitory activities against the indicator strain Lactobacillus sakei subsp. sakei JCM 1157(T). These results suggest that W. cibaria and W. paramesenteroides are the main LAB present during the fermentation of yan-dong-gua. This is the first report describing the distribution and varieties of LAB that exist in the yan-dong-gua fermentation process.

Characteristics of bacteriocin-like inhibitory substances from dochi-isolated Enterococcus faecium D081821 and D081833

To characterize bacteriocin-like inhibitory substances (BLIS) from two dochi-isolated Enterococcus faecium. Enterococcus faecium D081821 and D081833 were isolated from dochi (a traditional fermented food in Taiwan) and found to produce BLIS with inhibitory activities against Listeria monocytogenes, Clostridium perfringens, and Bacillus cereus. Strains D081821 and D081833 showed different growth temperatures and their BLIS showed different sensitivities to heat, proteolytic enzymes, and antibacterial spectra. Both BLIS were collected, partially purified, and analysed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). SDS-PAGE showed that both partially purified BLIS were approximately 3.0 kDa in size. These results indicate that E. faecium D081821 and D081833 produce different BLIS with strong antibacterial actions against the tested pathogenic bacteria. The results of this study suggest that two different BLIS from dochi-isolated lactic acid bacteria have potential for use as food preservatives.

Isolation and characterization of lactic acid bacteria from suan-tsai (fermented mustard), a traditional fermented food in Taiwan

To isolate, characterize, and identify lactic acid bacteria (LAB) in suan-tsai (fermented mustard), a traditional fermented food in Taiwan. Suan-tsai samples were collected at five time points from a fixed fermenting bucket. Fifty cultures were isolated from suan-tsai samples, and isolates were divided into classes by phenotype and then into groups by restriction-fragment length polymorphism analysis and sequencing of 16S ribosomal DNA. Phenotypic and biochemical characteristics identified two different bacterial groups (A and B), and the results showed that Pediococcus pentosaceus was the most abundant LAB during the initial fermentation time. However, the more NaCl-tolerant species Tetragenococcus halophilus took the place of P. pentosaceus and became the most abundant LAB later. All isolates were grown in de Man, Rogosa, and Sharpe (MRS) broth containing 6% NaCl, but T. halophilus could grow only in MRS broth containing 10% NaCl. These results suggest that the LAB P. pentosaceus and T. halophilus play roles in the fermentation of suan-tsai. This is the first report describing the distribution and varieties of LAB that exist in the suan-tsai fermentation process.

Isolation and characterization of lactic acid bacteria from dochi (fermented black beans), a traditional fermented food in Taiwan

To isolate, characterize and identify lactic acid bacteria (LAB) in dochi (fermented black beans), a traditional fermented food in Taiwan. A total of 30 samples were collected from three different dochi producers and analysed after different periods of storage. Fifty-two cultures of LAB were isolated from dochi samples and the isolates were divided into classes by phenotype and then into groups by restriction fragment length polymorphism analysis and sequencing of 16S ribosomal DNA. Phenotypic and biochemical characteristics identified six different bacterial groups (A-F) and showed that the majority of the isolates were homofermentative LAB. Enterococcus faecium was the most abundant of the dochi-isolated LAB. All isolated LAB were able to grow in MRS broth containing 6% NaCl, but only Enterococcus, Pediococcus and Tetragenococcus species could grow in MRS broth containing 10% NaCl. Furthermore, antibacterial activities of isolates were determined, and four isolates showed inhibitory activities against the indicator strain Lactobacillus sakei JCM 1157(T). These results suggest that Ent. faecium is the main LAB present during the fermentation of dochi. This is the first report describing the distribution and varieties of LAB that exist in the dochi fermentation process.