Bacillus sp. AIR-5, a strain from soil, produced an extracellular maltopentaose-forming amylase from amylose and soluble starch. This bacterium produced 8.9 g/l of maltopentaose from 40 g/l of soluble starch in a batch fermentation and the maltopentaose made up 90 % of the maltooligosaccharides produced (from maltose to maltoheptaose). The culture supernatant was concentrated using a 30 K molecular weight cut-off membrane and purified by DEAE-Cellulose and Sephadex G-150 column chromatographies. The purified protein showed one band on a native-PAGE and its molecular mass was estimated as 250 kDa. The 250-kDa protein was composed of tetramers of a 63-kDa protein. the isoelectric point of the purified protein was pH 6.9, and the optimum temperature for the enzyme activity was $45^{\circ}C$. The enzyme was quickly inactivated above $55^{\circ}C$, and showed a maximum activity at pH 8.5 and over 90% stability between a pH of 6 to 10. The putative N-terminal amino acid sequence of AIR-5 amylase, ATINNGTLMQYFEWYVPNDG, showed a 96% sequence similarity with that of BLA, a general liquefying amylase.
Protective effect of skin by antioxidative dietary buchu (Chinese chives, Allium tuberosum Router), was evaluated in ICR mice fed diets containing 2% or 5% buchu for 12 months. Lipid peroxidation and protein oxidation in skin, with or without ultraviolet B (UVB) irradiation, activities of antioxidative enzymes, total glutathione concentrations, and non-soluble collagen contents were measured. Dietary buchu decreased significantly in TBARS and protein carbonyl levels in skin compared to the control group, and were lower in those fed 5% than 2% buchu diet group. ICR mice exhibited an age-dependent decrease in antioxidative enzyme activities and total glutathione concentrations on the control diet, but in the groups fed buchu diet the enzyme activities and glu-tathione concentrations remained at youthful levels for most of the study. SOD, glutathione peroxidase, and catalase activities as well as total glutathione concentrations increased with time in the skins of the mice fed buchu diets. Lipid peroxidation and protein oxidation provoked by UVB irradiation on ICR mice skin homogenates were also significantly inhibited by dietary buchu. The buchu diets also decreased the formation of non-soluble collagen in mice skin, compared to the control group. These results suggest that antioxidative components and sulfur-compounds in buchu may confer protective effect against oxidative stress resulting from aging and exposure to ultraviolet irradiation.
Cortez-Vega, William Renzo;Fonseca, Gustavo Graciano;Bagatini, Daniela Cardozo;Prentice, Carlos
Food Science of Animal Resources
/
v.37
no.2
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pp.162-167
/
2017
Functional and nutritional soluble proteins can be recovered from surimi (and surimi-like material) processing wastewater, reducing environmental problems and the cost of an irresponsible waste disposal. Recovered proteins may be added back at a low percentage to surimi products. The aim of this work was to evaluate the effect of the addition of soluble recovered proteins (RP), obtained from mechanically separated chicken meat surimi-like material (MSCM-SLM) processing wastewater by acidic pH-shifting, on the composition and texture of RP-MSCM-SLM, with RP contents of 0, 10, 20 and 30% (w/w) in the mixture. For that, proximate composition and gel properties were evaluated. The fat content of the MSCM-SLM was significantly reduced to 11.98% and protein increased to 83.64% (dry basis) after three washing cycles. The addition of 30% RP in the MSCM-SLM significantly augmented the protein content to 93.45% and reduced fat content from to 2.78%. On the other hand, the addition of RP was responsible for a drastic decrease in texture parameters, reaching 252.36 g, 185.23 g.cm, and 6.97 N for breaking force, gel strength and cutting strength, respectively, when 30% of RP was included in the MSCM-SLM. It was concluded that the obtained intermediary product (RP-MSCM-SLM) is a good option to applications in processed meat products where high texture parameters are dispensable, e.g., emulsified inlaid frankfurter-type sausages, but high protein content and low fat content desired.
Nitrogen compounds of Panax ginseng and their biological activities in plant and animal were reviewed. Major nitrogen combounds found in P. ginseng are free amino acids, Water soluble teins, insoluble proteins and peptides. Minor nitrogen compounds are dencichine. glycol)roteins. amines, alkaloides, methoxy or alkyl pyrazine derivatives. free nucleosides and nllrleir arid bases. 4-me- thymi-5-thiazoleethanol and pyroglutamic acid. The contents of total nitrogen and protein in root increased until 13 years old rvhich was the highest age tinder investigation. Soluble protein content increased With the root weight and was higher in xylem pith than cortex-epidermis indicating the rlosc relation with root growth. Arginine which covered 58% of total free amino aroids may serve as a storage nitrogen. Arginine seems to be changed into proline in rhizome, threonine in stem and again threoning and arginine in leaf. The greater the root weight the higher the polyaminc content. Polyamine stimulated the growth of root callus. Physiological roles of other minor nitrogen compounds are unknown although dencichine content is relatively high (0.5% d.w.). biochemical and pharmatological activities of some nitrogen compounds for animal were more investigated than physiological roll iota plant itself. Radiation and U.V. protective function (heat stable protein), insulin-like activity in lipogenesis and lipolysis (adenosine and pyroglutamic acid), depression of blood sugar content (glycopeptide). hemostatir and nellrotoxic activity (denrichine) and. sedative and hypnotic activity (4-methyl-5-thiazoleethilnol) are reported. Heat stable protein increased with root age. The traditional quality critsria appear to be well in accordance with biological activities of nitrogen compounds. Chemical stlldies of nitrogen compounds seem relatively rare, probably dole to difficulty of isolation, subsequently the investigations of biological activities are little.
An In vitro protein sulfation in the soluble fraction of rat brain was charaderized further by an improved method of alkaline hydrolysis and thin layer ceflulose electrophoresis TLE) The protein sulfation was carried out in a reaction system containing [35 S] 3'-phosphoadenosine-5'-phosphosulfate (PAPS), Tris-maleate buffer (pH 8), MgCI$_2$, and soluble proteins from rat brain. The sulfated proteins were precipitated by acetone and alkaline hydrolysis was performed to obtain sulfated amino acids. The hydrolysate was separated further by TLE and the separated residues were identified by fluorography. The Iluorography of one-dimensional The showed at least nine sulfated residues including tryosine-O-sulfate. The other spots were not identified yet positively. General properties of protein sulfotransferases (PST) using this method were re-examined such as effects of concentrations of PAPS, pH, incubation temperature and $Mg^2$+. These results suggest a possible occurrence of several PST corresponding to each sulfated residue in rat brain and that the sulfation can occur not only in tyrosine but also in other residues as well.
Changes in the level of metabolites in leaves and pods were examined with respect to the seed chemical composition in black soybean. There was no further increase in pod length after 42 days after flowering (DAF). Pod weight, however, persistently increase until 73 DAF, thereafter the weight was slightly lowered. The seed storage protein, however, increased drastically as the increasing rate of pod weight was lessened at 61 DAF. The accumulation of seed storage proteins was occurred conspicuously as the increasing rate of pod weight was slowed down. The chlorophyll content both in leaves and pods was drastically decreased after 50 DAF. The beginning of drastic reduction in chlorophyll content was occurred concomitantly with the reduction of soluble protein content in leaves. The sugar content in leaves showed similar tendency with chlorophyll and soluble protein content. The starch level in leaves, however, showed different changing pattern during seed development. The starch content in leaves was increased persistently until 66 DAF, thereafter the content was decreased drastically to about $55\%$ of maximal value at 66 DAF. Total phenolics content in leaves and the anthocyanins content in seeds were stable without noticeable increase until 66 DAF. The contents were increased dramatically after 66 DAF showing the synchronized pattern with the decrease in starch level in leaves. The levels of the selected metabolites in leaf and seed suggested that the accumulation of chemical components of black soybean seed is launched actively at 66 DAF. The profile of storage proteins was nearly completed at 61 DAF because there was no large difference in densitometric intensity among protein subunits after 61 DAF. In soybean, chemical maturation of seed begins around 61 to 66 DAF at which most metabolites in vegetative parts are decreased and remobilized into maturing seeds.
Twenty-four new born crossbred (Bos indicus$\times$Bos taurus) calves were distributed in two equal groups and assigned to two different pre-starter diets with (Group 1) and without (Group 2) fish meal to study the effect of replacement of animal protein by vegetable protein in the diet and the age of animals on ruminal metabolic development. All calves were fed colostrum for 24 h and whole milk until weaning at 8 weeks of age. Rumen fluid samples were collected on 4 d, 1 wk, and then weekly interval up to 8 wk of age. Rumen fluid samples were analysed for pH, TVFA, lactic acid and N fractions (total N, total soluble N, trichloro acetic acid (TCA) soluble N, TCA precipitable N and ammonia N). Weekly feed intake and live weight gain pattern showed an increasing trend with the advancement of age, but were similar in both groups. The pH fell steadily during 0-4 wk of age and then stabilized in later period. A close relationship (r=0.80) between starter intake and TVFA concentration was observed in both the groups. Lactic acid (meq/l) and ammonia N (mg/dl) concentration showed initial rise (0.55 and 14.97 on day 4 to 3.38 (7 wk) and 32.85 (4 wk), respectively) to fall (2.74 and 17.60) again during 8 wk of age in response to increase in dry feed consumption (10% initially to 83% of diet dry matter at 8 wk of age). The TCA precipitable fraction of N did not show any change during 0-8 wk of age. Data indicate that the metabolic changes responded rapidly to dry feed intake which did not differ in fish meal and non-fish meal groups, and a poor voluntary consumption of oat hay retards the progressive changes in live weight and rumen microbial development.
Han, Sang Mi;Kim, Jung Min;Hong, In Phyo;Woo, Soon Ok;Kim, Se Gun;Jang, Hye Ri;Park, Kwan Kyu;Pak, Sok Cheon
Food Science of Animal Resources
/
v.35
no.5
/
pp.707-713
/
2015
Royal jelly has been widely used as a health supplement worldwide. However, royal jelly has been implicated in allergic reactions, and we developed a water-soluble royal jelly (WSRJ) without the allergy inducing protein. In this study, we aimed to identify the anti-melanogenic efficacy of WSRJ. B16F1 melanoma cells were first treated with 10 nM α-melanocyte stimulating hormone (α-MSH) and then with various doses of WSRJ. In addition, we investigated the mRNA and protein expression of melanogenesis-related genes such as tyrosinase, tyrosinase related protein-1 (TRP-1) and TRP-2 by reverse transcription-polymerase chain reaction and western blotting. WSRJ directly inhibited tyrosinase and cellular tyrosinase activity, which decreased melanin synthesis in α-MSH stimulated B16F1 melanoma cells a level comparable to that observed with arbutin. WSRJ decreased the mRNA and protein expressions of tyrosinase, TRP-1, and TRP-2, which was comparable to that observed with arbutin. WSRJ has strong anti-melanogenic activity, which invoice direct inhibition of tyrosinase enzyme activity and suppression of expression of melanogenesis related genes. Results from this study suggests that WSRJ is a potential candidate for the treatment of skin pigmentation.
The expression of the Paenibacillus sp. A11 cyclodextrinase (CDase) gene using the pUC 18 vector in Escherichia coli JM 109 resulted in the formation of an insoluble CDase protein in the cell debris in addition to a soluble CDase protein in the cytoplasm. Unlike the expression in Paenibacillus sp. A11, CDase was primarily observed in cytoplasm. However, by adding 0.5 M sorbitol as an osmolyte, the formation of insoluble CDase was prevented while a three-fold increase in cytoplasmic CDase activity was achieved after a 24 h-induction. The recombinant CDase protein was purified to approximately 14-fold with a 31% recovery to a specific activity of 141 units/mg protein by 40-60% ammonium sulfate precipitation, DEAE-Toyopearl 650 M, and Phenyl Sepharose CL-4B chromatography. It was homogeneous by non-denaturing and SDS-PAGE. The enzyme was a single polypeptide with a molecular weight of 80 kDa, as determined by gel filtration and SDS-PAGE. It showed the highest activity at pH 7.0 and $40^{\circ}C$. The catalytic efficiency ($k_{cat}/K_m$) values for $\alpha$-, $\beta$-, and $\gamma$-CD were $3.0{\times}10^5$, $8.8{\times}10^5$, and $5.5{\times}10^5\;M^{-1}\;min^{-1}$, respectively. The enzyme hydrolyzed CDs and linear maltooligosaccharides to yield maltose and glucose with less amounts of maltotriose and maltotetraose. The rates of hydrolysis for polysaccharides, soluble starch, and pullulan were very low. The cloned CDase was strongly inactivated by N-bromosuccinimide and diethylpyrocarbonate, but activated by dithiothreitol. A comparison of the biochemical properties of the CDases from Paenibacillus sp. A11 and E. coli transformant (pJK 555) indicates that they were almost identical.
This study was conducted to investigate the effects of the addition of varied levels of NaCI and phosphates to the breast meat of spent layers(2 \pm 0.2 kg), which were stabilized for over 24 h before slaughter, on the protein extractability, thiobarbituric acid(TBA) and volatile basic nitrogen (VBN). Within 1 h after slaughter, breast meat was removed and treated with NaCI(0, 1, 2, 3%) and phosphates(0.25% and 0.5%) using a hot-salted method. The breast meat samples were stored at 4\pm$1^{\circ}C$ for 3 d. The results obtained were summarized as follows. 1. Soluble protein contents of salt-treated groups were significantly higher than that of control (P<0.05) and showed a positive relationship with the levels of salt. At a constant level of NaCI, the soluble protein content was proportionately elevated by the levels of phosphates (P<0.05). It decreased significantly in both control and salt-treated groups during storage (P<0.05). 2. TBA values of salt-treated groups were significantly higher than that of control(P<0.05) and showed a positive relationship with the levels of salt. At a constant level of NaCI, TBA values in 0.5% phosphates treatment groups were significantly lower than that in 0.25%(P<0.05). It increased significantly in both control and salt-treated groups during storage(P<0.05). 3. VBN values of salt-treated groups were significantly lower than that of control(P<0.05) They increased significantly by the salt treatment for the first day of storage(P<0.05), but not from the second day of storage. VBN values in both control and salt-treated groups were significantly increased during storage(P<0.05). After the first day of storage and at the same level of NaCI, no significant difference in VBN value was observed between the two levels of phosphates.
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