• Applied Biological Chemistry
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• v.11
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• pp.1-27
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• 1969

More than thirty kinds of sea food pickles have been eaten in Korea. Out of these salted yellow tail pickle, salted clam pickle, salted oyster pickle, and salted cuttlefish pickle were employed for the analysis of their components, identification of main fermenting microbes, and determination of enzyme characteristics concerned. Also studied was the effect of enzymic action of microbes, which are concerned with the fermenting of pickles, on the production of flavorous 5'-mononucleotides and amino acids. The results are summarized as follows: 1. Microflora observed in the pickles are: (a) Total count of viable cells after 1-2 months of pickling was found to be $10^7$ and that after 6 months decreased to $10^4$. (b) Microbial occurence in the early stage of pickling was observed to be 10-20% Micrococcus spp., 10-20% Brevibacterium spp., 0-30% Sarcina spp., 20-30% Leuconostoc spp., ca 30% Bacillus spp., 0-10% Pseudomonas spp., 0-10% Flavobacterium spp., and 0-20% yeast. (c) Following the early stage of pickling, mainly halophilic bacteria such as Bacillus subtilis, Leuconostoc mesenteroides, Pediococcus halophilus and Sarcina litoralis, were found to exhibit an effect on the fermentation of pickle and their enzyme activities were in direct concern in fermentation of pickles. (d) Among the bacteria participating in the fermentation, Sarcina litoralis 8-14 and 8-16 strains were in need of high nutritional requirement and the former was grown only in the presence of purine, pyrimidine and cystine and the latter purine, pyrimidine and glutamic acid. 2. Enzyme characteristics studied in relation to the raw materials and the concerned microbes isolated are as follows: (a) A small amount of protease was found in the raw materials and 30-60% decrease in protease activity was demonstrated at 7% salt concentration. (b) Protease activity of halophilic bacteria, Bacillus subtilis 7-6, 11-1, 3-6 and 9-4 strains, in the complete media decreased by 10-30% at the 7% salt concentration and that of Sarcina litoralis 8-14 and 8-16 strains decreased by 10-20%. (c) Proteins in the raw materials were found to be hydrolyzed to yield free amino acids by protease in the fermenting microbes. (d) No accumulation of flavorous 5'-mononucleotides was demonstrated because RNA-depolymerase in the raw materials and the pickles tended to decompose RNA into nucleoside and phosphoric acid. (e) The enzyme produced in Bacillus subtilis 3-6 strain isolated from the salted clam pickles, was ascertained to be 5'-phosphodiesterase because of its ability to decompose RNA and thus accumulating 5'-mononucleotide. (f) It was demonstrated that the activity of phosphodiesterase in Bacillus subtilis 3-6 strain was enhanced by some components in the corn steep liquor and salted clam pickle. The enzyme activity was found to decrease by 10-30% and 40-60% at the salt concentration of 10% and 20%, respectively. 3. Quantitative data for free amino acids in the pickles are as follows: (a) Amounts of acidic amino acids such as glutamic and aspartic acids in salted clam pickle, were observed to be 2-10 times other pickles and it is considered that the abundance in these amino acids may contribute significantly to the specific flavor of this food. (b) Large amounts of basic amino acids such as arginine and histidine were found to occur in salted yellow tail pickle. (c) It is much interesting that in the salted cuttlefish pickle the contents of sulfur-containing amino acids were exceedingly high compared with those of others: cystine was found to be 17-130 times and methionine, 7-19 times. (d) In the salted oyster pickle a high content of some essential amino acids such as lysine, threonine, isoleucine and leucine, was demonstrated and a specific flavor of the pickle was ascribed to the sweet amino acids. Contents of alanine and glycine in the salted oyster pickle were 4 and 3-14 times as much as those of the others respectively. 4. Analytical data for 5'-mononucleotides in the pickles are as follows: (a) 5'-Adenylic acid and 3'-adenylic acid were found in large amounts in the salted yellow tail pickle and 5'-inosinic acid in lesser amount. (b) 5'-Adenylic acid, especially 3'-adenylic acid predominated in amount in the salted oyster pickle over that in the other pickles. (c) The salted cuttlefish pickle was found to contain only 5'-adenylic acid and 3'-adenylic acid. It has become evident from the above fact that clam and the invertebrate lack of adenylic deaminase and contain high content of adenylic acid. Thus, they were demonstrated to be the AMP-type. (d) 5'-Inosinic acid was contained in the salted yellow tail pickle in a significant concentration, and it might be considered to be IMP-type. 5. Comparative data for flavor with regard to the flavorous amino acids and the contents of 5'-mononucleotides are: (a) A specific flavor of salted yellow tail pickle was ascribed to the abundance in glutamic acid and aspartic acid, and to the existence of a small amount of flavorous 5'-inosinic acid. The combined effect of these components was belived to exhibit a synergistic action in producing a specific fiavor to the pickle. (b) A specific flavor of salted clam pickle has been demonstrated to be attributable to the richness in glutamic acid and aspartic acid rather than to that of 5'-mononucleotides.

• Journal of Korean Society of Forest Science
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• v.16 no.1
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• pp.1-32
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• 1972

By means of the interspecific hybridization in the Sub-genus Diploxylon of the Genus Pinus, $F_1$ hybrids of Pinus rigida${\times}$elliottii, Pinus rigida${\times}$radiata, P. rigida${\times}$serotina and P. densiflora${\times}$thunbergii had been produced. And on the basis of the crossabilities of these hybrids the taxonomic affinities of these pines were examined. And the needle characteristics of these hybrid and the occurence of phenolic substances in these $F_1$ hybrid were also investigated to see the potential usefulness of these characteristics for the diagnosis of the taxonomic affinity. And, the growth performances of the $F_1$ hybrids have also been compared with those of parental species. In order to contribute to the establishment of the hybrid seed orchard the introgression phenomena between P. densiflora and P. thunbergii in the eastern coastal area have also been investigated along with the investigation of the heterozygosity of plus trees of P. densiflora growing in the clone bank in Suwon. And the results were summarized as follows. 1. On the basis of crossabilities as well as on the taxonomic affinities according to the systems of Shaw, Pilger and Duffield, it has been proven that the parental species of those hybrids are of close affinities and range of the fertile hybrid seed production rate was as high as 28-58% in the best hybrid combination (Table 13). 2. Among those hybrids, the ${\times}$ Pinus, rigiserotina hybrid seemed to be most promising in the growth performance exhibiting 109-155% more volume growth compared to the seed parent with the statistic significance of 1% level (Tables 16 and 17). 3. Notwithstanding the fact that the all of the pollen parents are cold tender, all hybrids exhibit cold hardiness as much as their seed parent and it seems to suggest that the characteristics of cold hardiness were transmitted from the seed parent. 4. Though a striking difference in needle length was observed between the parental species of each hybrid, it was difficult to distinguish each hybrid from their seed parent by the needle length except ${\times}$P. rigiserotina which is characterized by long needle which is 65% more longer than the needle of the seed parent (Table 21). 5. With regard to the anatomical characteristics of needle, the hypoderm is apparently thicker in most of the $F_1$ hybrid pines and the characteristics of resin canals are dominated by medial in most $F_1$ hybrid. And, the fibrovascular bundles were apart as were in their seed parent. Therefore it was found to be possible to distinguish the hybrids pines from their parents by the needle characteristics. And, it is to be noticed that the ${\times}$P. densithunbergii was more close to the pollen parent having RDI value of 0.73 (Fig.l, Table 22). 6. It has been demonstrated that ${\times}$P. rigielliottii, ${\times}$P. rigiradiata and ${\times}$P. rigitaeda have a phenolic substance (No.7) of light yellow at Rf-0.46, same as their seed parent, but no trace of phenolic substance was observed in their pollen parent. This fact will serve as an important criteria for early identification of hybridity in progeny testing. However, the fact that both of ${\times}$P. rigiserotina and ${\times}$P. densithunbergii exhibit the same reactions of phenolic substances as well their parental species seems to indicate the close affinities between the parental species of the respective hybrid (Fig.2, Table 23). 7. The separation and the reaction of phenolic substance developed on TLC were found to be same in the same species showing no variations between the individuals, and no variations due to tree part of sampling, tree age or pollen sources. And the reaction was also observed regardless of the not varied by the kind of developing solvent whether it is Aceton-Chloroform (3:7 v/v) or Benzene-Methanol-Acetic acid (90:16:8 v/v). 8. The introgression phenomena of natural Pinus densifiora stand in both east and west coastal area indicates that the major part of the red pines investigated are all heterozygous and the heterozygosity of pines are higher in the west coast than in the east coast(Tables 24 and 25). 9. Based on the RDI, among the plus trees of Pinus densiflora selected in Korea and Japan as well, no pure P. densiflora has been found. Since all of the sample trees of Pinus densiflora were found to be as heterozygous bearing part of the characteristics of P. thunbergii, those red pines were considered to be natural heterotic hybrid pines(Figs. 3 and 4. Tables 26 and 27).