• Journal of the Korean Society of Food Science and Nutrition
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• v.30 no.5
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• pp.782-786
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• 2001

The taste characteristics and functionality of low-utilized small longfinned squid as affected by two stage enzyme hydrolysis were examined. In taste active-components, total free amino acid contents in hot-water and autolytic extract, two stage enzyme hydrolysate (TSEH) of longfinned squid were 2,792.5 mg%, 8,393.8 mg% and 9,186.1 mg%, respectively. The major free amino acids were Pro, Leu, Glu, Tau, Lys, Arg, Phe, Val and Ile. As for quarternary ammonium bases, betaine was the principal component (593.8 mg%) and also contents of TMAO, AMP in longfinned squid TSEH were 234.8% mg% and 51.0 mg%, respectively. The major inorganic ions in TSEH were Na(874.0 mg%), K (398.2 mg%), Cl (1,213.1 mg%) and PO$_4$(995.9 mg%). From the results in sensory tests, TSEH was superior to other extracts on the aspects of taste characteristics such as umami intensity, sweetness, taste harmony and transparency of extract. Also TSEH of longfinned squid revealed very higher Angiotensin-I converting enzyme inhibition ratio (92.1%) than those of hot-water and autolytic extract.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.36 no.3
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• pp.210-219
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• 2003

To develop natural flavoring substances, optimal two stage enzyme hydrolysis conditions and flavor compounds of short-neck clam (Tapes philippinarum) enzyme hydrolysates were examined. The optimal enzyme hydrolysis conditions for two stage enzyme hydrolysate (TSEH) of short-neck clam were revealed in temperature at $55^{\circ}C$ for 4 hours digestion with alcalase at the 1st stage and 4 hours digestion at $45^{\circ}C$ with exopeptidase type neutrase at the 2nd stage. In quality tests of hot-water extracts, steam extracts and 4 kinds of enzyme hydrolysates, TSEH processing method was superior to other methods in yield, nitrogen contents, organoleptic taste such as umami intensity and inhibition of off-flavor formation, and transparency of extract. Total free amino acid contents in hot-water extract, steam extract and TSEH were 1,352.1 mg/100 g, 1,174.1 mg/100 g and 2,122.4 mg/100 g, respectively, Major free amino acids in TSEH were glutamic acid, glycine, alanine, tyrosine, phenylalanine and arginine. As for nucleotides and other bases, betaine, TMAO and creatinine were principal components in TSEH. The major inorganic ions in TSEH were Na, K, P and Cl. TSEH also revealed very higher angiotensin-I converting enzyme inhibition effect $(70.7\%)$ than those of hot-water and steam extract. We conclude that TSEH from short-neck clam was more flavorable compared with the seasoning materials on the market, it could be utilized as the instant soup base and the seasoning substances for fisheries processing.

• Asian-Australasian Journal of Animal Sciences
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• v.16 no.9
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• pp.1384-1389
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• 2003

In order to clarify one of the biological functions of pork, we investigated whether a peptic hydrolysate of denatured porcine crude myosin showed inhibitory activity against angiotensin I-converting enzyme (ACE), which contributed to hypertension. Our results indicated that this hydrolysate showed relatively strong activity, and we therefore attempted to separate the involved peptides, which were considered to be active substances. To isolate these active peptides, the hydrolysate was separated using a solidphase separation, gel filtration high-performance liquid chromatography (HPLC), and two kinds of reverse phase HPLC. In each stage of separation, many fractions were detected, almost all of which showed ACE inhibitory activity. Thus, we suggested that the activity of the hydrolysate as a whole was a result of the activities of the many individual peptides. Six peaks were distinguished, with yields from 34 to 596 ppm of original crude myosin. In addition to the six peaks, many other active fractions were found throughout the separation steps, strongly suggesting that whole porcine crude myosin itself had ACE inhibitory activity. Moreover, pork as food was considered to function as an ACE inhibitory material in vivo, because pork proteins consist primarily of crude myosin, which included almost all the myofibrillar structural proteins.

• Applied Biological Chemistry
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• v.37 no.2
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• pp.130-141
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• 1994

A continuous two-stage membrane (1st-SCMR, MWCO 10,000; 2nd-SCMR, MWCO 5,000) reactor was developed and optimized for the production of fish skin gelatin hydrolysate with different molecular size distribution profiles using trypsin and pronase E. The optimum operating conditions in the 1st-step membrane reactor using trypsin were: temperature, $55^{\circ}C$ ; pH 9.0; enzyme concentration, 0.1 mg/ml; flux, 6.14 ml/min; reaction volume, 600 ml; and the ratio of substrate to trypsin, 100 (w/w). After operating for 1 hr under the above conditions, 79% of total amount of initial gelatin was hydrolysed. In the 2nd-step using pronase E under optimum operating conditions[temperature, $50^{\circ}C$ ; pH 8.0; enzyme concentration, 0.3 mg/ml; flux, 6.14 ml/min; reaction volume, 600 ml; and the ratio of substrate to pronase E, 33 (w/w)], the 1st-step hydrolysate was hydrolysed above 80%. Total enzyme leakages in the 1st-step and 2nd-step membrane reactors were about 11.5% at $55^{\circ}C$ for 5hrs and 9.0% at $50^{\circ}C$ for 4 hrs, respectively. However, there was no apparent correlation between enzyme leakage and substrate hydrolysis. The membrane has a significant effect on activity lose of trypsin and pronase E activity for 1 hr of the membrane reactors operation. The loss of initial activity of enzymes were 34% and 18% in the 1st-step and 2nd-step membrane reactor, whereas were 23% and 10% after operating time 3 hr in the 1st-step and 2nd-step membrane reactor lacking the membrane, respectively. The productivities of 1st-step and 2nd-step membrane reactor for 8 times of volume replacement were 334 mg and 250 mg per mg enzyme, respectively.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.35 no.3
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• pp.259-264
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• 2002

In order to develope nutritional and flavoring intermediate products, the optimal processing conditions for two stage enzyme hydrolysate (TSEH) from low-utilized conger eel scrap such as head and intestine were investigated. The optimal processing conditions for TSEH were revealed in temperature at $55^{\circ}C$ 3$\~$4 hours digestion with alcalase at the 1st stage, and 4 hours at $45{\~}50^{\circ}C$ digestion with neutrase at the 2nd stage. Among water extract, steam extract and enzyme hydrolysates of conger eel scrap, the present TSEH was superior to other extracts in terms of yield ana organoleptic taste such as harmonic umami and inhibition of fishy and greasy taste formation. From the results of chemical experiments and sensory evaluation, we may conclude that TSEH of conger eel scrap could be utilized as the flavoring intermediate materials for the fisheries products such as flavoring sauces, drinkable beverage and instant food materials.

• Journal of the Korean Society of Food Science and Nutrition
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• v.29 no.4
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• pp.663-668
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• 2000

To develop natural intermediate flavoring substances, optimal processing conditions and qualities for two stage enzyme hydrolysate (TSEH) from low-utilized small longfinned squid were investigated. The optimal conditions for TSEH method were found as digestion with Alcalase (0.2% w/w-sample, pH 8.0) at 55$^{\circ}C$ 3 hours at the 1st stage and with Neutrase (exo-peptidase, 0.2% w/w-sample, pH 6.0) at 45$^{\circ}C$ for 2~3 hours at the 2nd stage. Among the method of water extract, autolytic extract and various kinds yields, transparency and organoleptic taste. From the results of chemical experiments and sensory evaluation, longfinned squid TSEH is flavorable as the natural intermediate taste-active substances for fisheries products such as soup base, squid-taste pasty and snacks.

• Korean Journal of Food Science and Technology
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• v.30 no.6
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• pp.1339-1344
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• 1998

To develop natural flavoring substances, optimal hydrolysis conditions for two stage enzyme hydrolysates (TSEH) using small kingfish (Maegari) were investigated. The optimal conditions for TSEH were revealed in temperature at $50^{\circ}C$ for 3 hours digestion with alcalase (Aroase AP-10, pH 8.0) at the 1st stage and 2 hours digestion at $45^{\circ}C$ with neutrase (Pandidase NP-2, pH 6.0) at the 2nd stage. From the results in quality tests of water extracts, autolytic extracts and 4 kinds of enzyme hydrolysates, TSEH processing method was superior to other methods on the aspects of yield, nitrogen contents, taste such as umami intensity and inhibition of off-flavor formation, and transparency of extracts. We may conclude that TSEH from small kingfish was more flavorable compared with the conventional seasoning materials, it could be utilized as the seasoning substances for fisheries processing.

• Journal of Microbiology and Biotechnology
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• v.25 no.2
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• pp.185-195
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• 2015

Glucoamylase is an important industrial enzyme. Glucoamylase production by industrial Aspergillus niger strain featured with two major problems: (i) empirical substrate feeding methods deteriorating the fermentation performance; and (ii) the high raw materials cost limiting the economics of the glucoamylase product with delegated specification. In this study, we first proposed a novel three-stage varied-rate substrate feeding strategy for efficient glucoamylase production in a 5 L bioreactor using the standard feeding medium, by comparing the changing patterns of the important physiological parameters such as DO, OUR, RQ, etc., when using different substrate feeding strategies. With this strategy, the glucoamylase activity and productivity reached higher levels of 11,000 U/ml and 84.6 U/ml/h, respectively. The performance enhancement in this case was beneficial from the following results: DO and OUR could be controlled at the higher levels (30%, 43.83 mmol/l/h), while RQ was maintained at a stable/lower level of 0.60 simultaneously throughout the fed-batch phase. Based on this three-stage varied-rate substrate feeding strategy, we further evaluated the economics of using alternative carbon sources, attempting to reduce the raw materials cost. The results revealed that cornstarch hydrolysate could be considered as the best carbon source to replace the standard and expensive feeding medium. In this case, the production cost of the glucoamylase with delegated specification (5,000 U/ml) could be saved by more than 61% while the product quality be ensured simultaneously. The proposed strategy showed application potential in improving the economics of industrial glucoamylase production.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.31 no.6
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• pp.791-798
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• 1998

To develop natural flavoring substances. optimal hydrolysis conditions for two stage enzyme hydrolysates (TSEH) using low-utilized shellfishes such as purplish clam and frozen oyster stored at $-20^{\circ}C$ for 60 days. The optimal conditions for TSEH method were revealed in temperature at $50^{\circ}C$ 3 hours digestion with alcalase (Aroase AP-10, $0.3%$ w/v, pH 8.0) at the 1st stage and $45^{\circ}C$ 2 hours digestion with neutrase (Pandidase NP-2, $0.3\%$ w/v, pH 6.0) at the 2nd stage. Among water extracts, autolytic extracts and 4 kinds of enzyme hydrolysates tests, TSEH method was superior to other methods on the aspect of yields, nitrogen contents, taste such as umami and control of off-flayer formation, and transparency of extracts. From the results of chemical experiments and sensory evaluation, we may conclude that TSEH from low-utilized marine products is more flavorable compared the conventional enzyme hydrolysates, it could be commercialized as the seasoning substances.

• Korean Journal of Food Science and Technology
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• v.30 no.6
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• pp.1345-1349
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• 1998

The flavor constituents and functionality of small kingfish were studied as affected by two stage enzyme hydrolysis (TSEH). Total free amino acid contents in water extract, autolytic extract and TSEH of small kingfish were 541.3 mg%, 8,245.3 mg% and 6,636.6 mg%, respectively. Major free amino acids in TSEH were hydroxyproline, glutamic acid, proline, leucine, phenylalanine, lysine, arginine. As for nucleotides and other bases, IMP, TMAO and total creatinine were principal components in TSEH. And the major inorganic ions in TSEH were Na, K, P, Cl and $PO_{4}$. Also TSEH of small kingfish revealed very higher Angiotensin-I converting enzyme inhibition effect than those of water and autolytic extract.