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Determination of Microbial Diversity in Gouda Cheese via Pyrosequencing Analysis
Oh, Sangnam,Kim, Younghoon Korean Society of Dairy Science and Biotechnology 2018 한국유가공기술과학회지 Vol.36 No.2
The present study aimed to investigate the microbial diversity in Gouda cheese within the four months of ripening, via next-generation sequencing (NGS). Lactococcus (96.03%), and Leuconostoc (3.83%), used as starter cultures, constituted the majority of bacteria upon 454 pyrosequencing based on 16S rDNA sequences. However, no drastic differences were observed among other populations between the center and the surface portions of Gouda cheese during ripening. Although the proportion of subdominant species was <1%, slight differences in bacterial populations were observed in both the center and the surface portions. Taken together, our results suggest that environmental and processing variables of cheese manufacturing including pasteurization, starter, ripening conditions are important factors influencing the bacterial diversity in cheese and they can be used to alter nutrient profiles and metabolism and the flavor during ripening.
Comparison of Total RNA Isolation Methods for Analysis of Immune-Related microRNAs in Market Milks
Sangnam Oh,Mi Ri Park,Seok Jun Son,Younghoon Kim 한국축산식품학회 2015 한국축산식품학회지 Vol.35 No.4
Bovine milk provides essential nutrients, including immunologically important molecules, as the primary source of nutrition to newborns. Recent studies showed that RNAs from bovine milk contain immune-related microRNAs (miRNA) that regulate various immune systems. To evaluate the biological and immunological activity of miRNAs from milk products, isolation methods need to be established. Six methods for extracting total RNAs from bovine colostrums were adopted to evaluate the isolating efficiency and expression of miRNAs. Total RNA from milk was presented in formulation of small RNAs, rather than ribosomal RNAs. Column-combined phenol isolating methods showed high recovery of total RNAs, especially the commercial columns for biofluid samples, which demonstrated outstanding efficiency for recovering miRNAs. We also evaluated the quantity of five immune-related miRNAs (miR-93, miR-106a, miR-155, miR-181a, miR-451) in milk processed by temperature treatments including low temperature for long time (LTLT, 63oC for 30 min)-, high temperature for short time (HTST, 75oC for 15 s)-, and ultra heat treatment (UHT, 120-130oC for 0.5-4 s). All targeted miRNAs had significantly reduced levels in processed milks compared to colostrum and raw mature milk. Interestingly, the amount of immune-related miRNAs from HTST milk was more resistant than those of LTLT and UHT milks. Our present study examined defined methods of RNA isolation and quantification of immune-specific miRNAs from small volumes of milk for use in further analysis.
Comparison of Total RNA Isolation Methods for Analysis of Immune-Related microRNAs in Market Milks
Oh, Sangnam,Park, Mi Ri,Son, Seok Jun,Kim, Younghoon Korean Society for Food Science of Animal Resource 2015 한국축산식품학회지 Vol.35 No.4
Bovine milk provides essential nutrients, including immunologically important molecules, as the primary source of nutrition to newborns. Recent studies showed that RNAs from bovine milk contain immune-related microRNAs (miRNA) that regulate various immune systems. To evaluate the biological and immunological activity of miRNAs from milk products, isolation methods need to be established. Six methods for extracting total RNAs from bovine colostrums were adopted to evaluate the isolating efficiency and expression of miRNAs. Total RNA from milk was presented in formulation of small RNAs, rather than ribosomal RNAs. Column-combined phenol isolating methods showed high recovery of total RNAs, especially the commercial columns for biofluid samples, which demonstrated outstanding efficiency for recovering miRNAs. We also evaluated the quantity of five immune-related miRNAs (miR-93, miR-106a, miR-155, miR-181a, miR-451) in milk processed by temperature treatments including low temperature for long time (LTLT, 63℃ for 30 min)-, high temperature for short time (HTST, 75℃ for 15 s)-, and ultra heat treatment (UHT, 120-130℃ for 0.5-4 s). All targeted miRNAs had significantly reduced levels in processed milks compared to colostrum and raw mature milk. Interestingly, the amount of immune-related miRNAs from HTST milk was more resistant than those of LTLT and UHT milks. Our present study examined defined methods of RNA isolation and quantification of immune-specific miRNAs from small volumes of milk for use in further analysis.
Determination of Microbial Diversity in Gouda Cheese via Pyrosequencing Analysis
( Sangnam Oh ),( Younghoon Kim ) 한국유가공기술과학회 2018 한국유가공기술과학회지 Vol.36 No.2
The present study aimed to investigate the microbial diversity in Gouda cheese within the four months of ripening, via next-generation sequencing (NGS). Lactococcus (96.03%), and Leuconostoc (3.83%), used as starter cultures, constituted the majority of bacteria upon 454 pyrosequencing based on 16S rDNA sequences. However, no drastic differences were observed among other populations between the center and the surface portions of Gouda cheese during ripening. Although the proportion of subdominant species was <1%, slight differences in bacterial populations were observed in both the center and the surface portions. Taken together, our results suggest that environmental and processing variables of cheese manufacturing including pasteurization, starter, ripening conditions are important factors influencing the bacterial diversity in cheese and they can be used to alter nutrient profiles and metabolism and the flavor during ripening.
( Sangnam Oh ),( Mi-ri Park ),( Sangdon Ryu ),( Brighton E. Maburutse ),( Ji-uk Kim ),( Younghoon Kim ) 한국미생물생명공학회(구 한국산업미생물학회) 2017 Journal of microbiology and biotechnology Vol.27 No.9
MicroRNAs (miRNAs) are abundant in bovine milk and milk derived from other livestock, and they have functional roles in infants and in the secretion process of mammary glands. However, few studies have evaluated miRNAs in dairy processes, such as during cheese making and ripening. Thus, we investigated the characteristics of milk-derived miRNAs during the manufacturing and ripening of Camembert cheese as well as the microbiota present using the quantitative reverse transcription polymer chain reaction (RT-qPCR) and 16S rRNA pyrosequencing, respectively. Pyrosequencing showed that the cheese microbiota changed dramatically during cheese processing, including during the pasteurization, starter culture, and ripening stages. Our results indicated that the RNA contents per 200 mg/200 μl of the sample increased significantly during cheese-making and ripening. The inner cheese fractions had higher RNA contents than the surfaces after 12 and 22 days of ripening in a timedependent manner (21.9 and 13.2 times higher in the inner and surface fractions than raw milk, respectively). We performed a comparative analysis of the miRNAs in each fraction by RT-qPCR. Large amounts of miRNAs (miR-93, miR-106a, miR-130, miR-155, miR-181a, and miR- 223) correlated with immune responses and mammary glands were present in aged cheese, with the exception of miR-223, which was not present on the surface. Considerable amounts of miRNAs were also detected in whey, which is usually disposed of during the cheese-making process. Unexpectedly, there were no significant correlations between immune-related miRNAs and the microbial populations during cheese processing. Taken together, these results show that various functional miRNAs are present in cheese during its manufacture and that they are dramatically increased in amount in ripened Camembert cheese, with differences according to depth.