• Journal of Microbiology and Biotechnology
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• v.21 no.6
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• pp.617-626
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• 2011

Transglutaminase from Streptomyces mobaraensis is an enzyme of unknown function that cross-links proteins to high molecular weight aggregates. Previously, we characterized two intrinsic transglutaminase substrates with inactivating activities against subtilisin and dispase. This report now describes a novel substrate that inhibits papain, bromelain, and trypsin. Papain was the most sensitive protease; thus, the protein was designated Streptomyces papain inhibitor (SPI). To avoid transglutaminase-mediated glutamine deamidation during culture, SPI was produced by Streptomyces mobaraensis at various growth temperatures. The best results were achieved by culturing for 30-50 h at $42^{\circ}C$, which yielded high SPI concentrations and negligibly small amounts of mature transglutaminase. Transglutaminasespecific biotinylation displayed largely unmodified glutamine and lysine residues. In contrast, purified SPI from the $28^{\circ}C$ culture lost the potential to be cross-linked, but exhibited higher inhibitory activity as indicated by a significantly lower $K_i$ (60 nM vs. 140 nM). Despite similarities in molecular mass (12 kDa) and high thermostability, SPI exhibits clear differences in comparison with all members of the wellknown family of Streptomyces subtilisin inhibitors. The neutral protein (pI of 7.3) shares sequence homology with a putative protein from Streptomyces lavendulae, whose conformation is most likely stabilized by two disulfide bridges. However, cysteine residues are not localized in the typical regions of subtilisin inhibitors. SPI and the formerly characterized dispase-inactivating substrate are unique proteins of distinct Streptomycetes such as Streptomyces mobaraensis. Along with the subtilisin inhibitory protein, they could play a crucial role in the defense of vulnerable protein layers that are solidified by transglutaminase.

• Food Science of Animal Resources
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• v.35 no.3
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• pp.360-369
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• 2015

Milk proteins have many potential sequences within their primary structure, each with a specific biological activity. In this study, we compared and investigated the bioactivities of hydrolysates of the domestic (A, B) and imported (C, D) skim milk powders generated using papain digestion. MALDI-TOF analysis revealed that all milk powder proteins were intact, indicating no autolysis. Electrophoretic analysis of hydrolysates showed papain treatment caused degradation of milk proteins into peptides of various size. The antioxidant activity of the hydrolysates, determined using 2,2-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and total phenolic contents (TPC) assays, increased with incubation times. In all skim milk powders, the antioxidant activities of hydrolysates were highest following 24 h papain treatment (TPC: A, 196.48 μM GE/L; B, 194.52 μM GE/L; C, 194.76 μM GE/L; D, 163.75 μM GE/L; ABTS: A, 75%; B, 72%; C, 72%; D, 57%). The number of peptide derived from skim milk powders, as determined by LC-MS/MS, was 308 for A, 283 for B, 208 for C, and 135 for D. Hydrolysate A had the highest antioxidant activity and the most potential antioxidant peptides amongst the four skim milk powder hydrolysates. A total of 4 β-lactoglobulin, 4 α_s1-casein, and 56 β-casein peptide fragments were identified as potential antioxidant peptides in hydrolysate A by LC-MS/MS. These results suggest that domestic skim milk could have applications in various industries, i.e., in the development of functional foods.

• Korean Journal of Food Science and Technology
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• v.6 no.3
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• pp.163-168
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• 1974

Authors investigated the tenderness effect of beef which was treated with papain and obtained results indicated as follow. 1. By using the sensory test, most desirable tenderness were obtained in 0.05% of enzyme solution for 40minute to 1 hour at $25^{\circ}C$. 2. It was also somewhat recognized as a tenderness effect with treated for 40minute to 1 hour at $10^{\circ}C$. 3. Water soluble and amino acid nitrogen were increased in proportion to enzyme concentration, reaction temperature and reaction time and opposite in connective tissue nitrogen. 4. It will be recognized that there were correlation between result of sensory test and water soluble, amino acid and connective tissue nitrogen.

• Journal of the Korean Society of Food Science and Nutrition
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• v.17 no.3
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• pp.233-241
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• 1988

In order to develop a new type of food source for the effective utilization of fish protein, plastein reaction was applied to improve the functional properties of sardine protein. Conditions necessary for optimal plastein productivity from sardine protein using pepsin, ${\alpha}-chymotrypsin$, protease(from Aspergillus saitoi) and papain were established. Sardine protein concentrate was hydrolyzed with pepsin yielding an approximate degree of hydrolysis of 78.4%. Enzyme induced plastein was optimized at : pH 4 for pepsin, pH 7 for ${\alpha}-chymotrypsin$, pH 5 for pretense and pH 6 for papain : Substrate concentrate 40% for pepsin and ${\alpha}-chymotrypsin$, 50% for pretense and papain : the time of incubation, 24hr : enzyme/substrate ratio, 1 : 100(w/v) incubation temperature, $50^{\circ}C$.

• Food Science of Animal Resources
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• v.38 no.1
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• pp.143-151
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• 2018

The effects of proteolytic enzymes (bromelain and bromelain+papain) and a ginger extract were assessed on collagen content and solubility, thermal shrinkage temperature of connective tissue, pH, cooking loss, drip loss, and Warner-Bratzler shear force (WBSF) of M. pectoralis profundus isolated from the beef brisket cut. Both proteolytic enzymes and ginger extract led to a significant increase in cooking loss and collagen solubility compared with untreated controls. On the other hand, the peak ($T_p$) thermal shrinkage temperature markedly decreased in all treatments compared with those in controls. Samples treated with bromelain, bromelain + papain, and ginger extract showed a significant decrease in WBSF by 36%, 40%, and 37%, respectively, compared with untreated controls. Our findings suggest that ginger extract are useful for post-mortem tenderization of meat containing high levels of collagen, compared to control even though, bromelain and bromelain + papain treatments have higher collagen solubility than ginger extract.

• Microbiology and Biotechnology Letters
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• v.18 no.5
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• pp.501-505
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• 1990

Streptomyces sp. KISl3 isolated from soil was found to produce low molecular weight thiol protease inhibitors. The protease inhibitor production was closely linked to the cell growth and regulated by growth condition. The inhibitor was purified from the culture broth through butanol extraction, silicagel 60 column chromatography, Sephadex LH-20 gel filtration and preparative HPLC. The inhibitor showed specific inhibitory activity to thiol protease such as papain, picin and bromelain. The mode of inhibition against papain to Hammersten casein as a substrate was non-competitive.

• The Korean Journal of Food And Nutrition
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• v.16 no.4
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• pp.379-387
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• 2003

The freeze dried young antler was extracted by water and proteases. In case of water extraction, the extraction rate was highest when it was reacted in 5% of concentration for 6 hours at 50$^{\circ}C$. The result of HPLC analysis of extract shows that high molecular peak in water extract was transformed into low molecular polk by proteases. The rate of low molecular peak was highest when bacteria protease was used, and its second highest rate was pepsin, but the effect of papain on it was low, The extraction rate of young antler reacted for 5 hours was 33.4%(absorbance 13.25 at 280nm) of bacteria protease, 22.4%(absorbance 10.06) of papain, and 30.2% (absorbance 11.34) of pepsin. The young antler was boiled for 30min and it was reacted by proteases for 5 hours at 50$^{\circ}C$. The extraction rate of it was 47,6%(absorbance 12,54) of bacteria protease, and 26,4%(absorbance 7,48) of papain, and 45.6%(absorbance 7.23) of pepsin, In protein content, water extract was 52,1%, bacteria protease extract was 37.8%, and in amino acid content, water extract was 16.3%, bacteria protease extract was 31.96%, in ash content, water extract was 8.8%, bacteria protease extract was 5.6% by dry base. In mineral content, water extract contains 3.6% of Ca, 8.6% of P, 0.01% of Mg, 1.4 % of Na, 0.02 % of F, and bacteria protease extract contains 2.5% of Ca, 11.8% of P, 0.046 % of Mg, 2.1 % of Na, 0.018 % of F by dry base.

• Korean Journal of Food Science and Technology
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• v.11 no.3
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• pp.171-175
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• 1979

Thermal properties of crude papain and crude peroxidase from domestic papaya were investigated. The crude extract of papaya was inactivated at the temperature range of $60^{\circ}{\sim}90^{\circ}C$ at pH 7.0 and the rest of the activities of papain and peroxidase were determined, respectively. The heat inactivation of papain and papaya peroxidase was biphasic at low temperature. For the thermal inactivation of papain extract, the enthalpy of activation was 91.4 kJ/mol, the entropy of activation, -49.6 J/mol K, and the free energy of activation, 108.5 kJ/mol. The activation energy for the inactivation of papaya peroxidase was 168.5 kJ/mol, the entropy of activation, $200.4\;J/mol{\cdot}K$ and the free energy of activation, 99.7 kJ/mol. The thermal stability of papain showed that it has a possibility for use as a meat tenderizer. It was also discussed that papaya peroxidase could be more suitable as a biochemical criteria for heat treatment than papaya catalase.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.32 no.4
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• pp.427-431
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• 1999

This study was conducted to investigate the inhibitory activity of water extracts and its enzymatic hydrolysates from algae against angiotensin-I converting enzyme (ACE). The 7 kinds of algae were extracted with water at $50^{\circ}C,\;70^{\circ}C$ and $98^{\circ}C$. ACE inhibitory activities of water extracts were the highest at $70^{\circ}C$, and those of ceylon moss, layer, green layer, sea mustard, seaweed fusiforme sea tangle and sea staghorn were $10.9\%,\;9.3\%,\;8.9\%,\;8.2\%,\;7.5\%,\;7.1\%$ and $7.0\%$, respectively. Layer, green laver sea mustard and ceylon moss of high ACE inhibitory activities among the 7 kinds of water extracts were hydrolyzed by maxazyme and papain during 24hrs. ACE inhibitory activity of enzymatic hydrolysates was higher than that of water extracts, and was the highest in enzymatic hydrolysates of laver among the tested samples. In laver hydrolysates by proteases, the highest ACE inhibitory activity and peptide-nitrogen contents were observed at 8 hours hydrolysis and the hydrolysates by maxazyme showed relatively higher activity than those by papain(31.3 and $27.9\%$, respectively). But peptide-nitrogen contents were greater in papain hydrolysates than in maxazyme.

• Journal of the Korean Wood Science and Technology
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• v.32 no.5
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• pp.39-44
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• 2004

This study was carried out to investigate the tenderizing effect of Neungi mushroom (Sarcodon aspratus) powder and its protease. The addition of Neungi mushroom powder and its protease enhanced water retention values (WRY) of meat. The WRY of meat was increased 26.8% by protease addition, compared to 13.8% WRV by sugar addition. This increase in WRY derived to the increase of water soluble fraction in the meat texture by hydrolysis of meat protein, and had the meat tenderized. Concerned to the meat tenderizing effect, the addition of Neungi mushroom powder and its protease have decreased of meat hardness and gave similar tenderizing effect, as compared to commercial tenderizer, papain. The decreasing rates of meat hardness were 51.6% of Neungi mushroom powder, 58.5% of its protease, and 563% of commercial tenderizer, papain. This tenderizing effect of protease attributed to the degradation of muscle fiber protein in meat, such as actin, myosin and connectin etc. The addition of Neungi mushroom to foods gives significant changes in food color, mainly decreasing lightness.