Comparative Analysis of the Elementary and Amino Acid Composition of Nepeta olgae Regel L. Plants Growing in Uzbekistan

This article for the first time presents the results of the study of qualitative and quantitative elemental and amino acid composition of the aboveground part of the plant Nepeta olgae Regel (L.) taken in the territory of Chust and Kosonsai districts (from the slopes of Gova and Kosonsai mountains) of Namangan region during the period before and during flowering (May-June, 2021-2022). The use of instrumental analysis of high-throughput energy dispersive X-ray fluorescence spectrometry, allowed to establish 20 mineral elements in the plant Nepeta olgae Regel (L.), among which to vital 9 elements and 3 to conditionally necessary. The amino acid composition of the plant Nepeta olgae Regel (L.) was studied by high performance liquid chromatography (HPLC) and 17 compounds were identified. Of these, 8 were substitutable and 9 essential amino acids.

Prediction of protein subcellular localization based on multilayer sparse coding / 生物工程学报

In order to provide a theoretical basis for better understanding the function and properties of proteins, we proposed a simple and effective feature extraction method for protein sequences to determine the subcellular localization of proteins. First, we introduced sparse coding combined with the information of amino acid composition to extract the feature values of protein sequences. Then the multilayer pooling integration was performed according to different sizes of dictionaries. Finally, the extracted feature values were sent into the support vector machine to test the effectiveness of our model. The success rates in data set ZD98, CH317 and Gram1253 were 95.9%, 93.4% and 94.7%, respectively as verified by the Jackknife test. Experiments showed that our method based on multilayer sparse coding can remarkably improve the accuracy of the prediction of protein subcellular localization.

System isolation and purification of Isatidis Radix polysaccharides and determination of their compositions / 中草药

Objective: To isolate and purify the polysaccharides (glycoproteins) from IsatidisRadix (Banlangen) systematically and to study the composition of them. Methods: Crude polysaccharides precipitated by 80% ethanol from thewater extract of IsatidisRadix, which has theanti-viral activity, were fractionated by DEAE-Sepharose Fast Flow, Sephacryl-200, and high gel chromatography system sequentially. The composition of monosaccharides and amino acids of polysaccharides (glycoproteins) was then determined by HPLC with pre-column derivatization using 1-phenyl-3-methyl-5-pyrazolone (PMP) and o-phthalaldehyde (OPA), respectively. Results: Two of homogeneous polysaccharides named IRPS1A and IRPS1B and two of homogeneous glycoproteins named IRPS2A and IRPS3A were obtained from IsatidisRadix by systematical separation and purification. The monosaccharide composition of IRPS1A and IRPS1B was of arabinose, mannose, galactose, and glucose. IRPS2A contained galacturonic acid, glucose, galactose, and arabinose. While IRPS3A contained mannose, rhamnose, galacturonic acid, glucose, galactose, and arabinose. The amino acid compositions of IRPS2A and IRPS3A were 14 kinds of amino acid residues including Asp, Glu, Ser, His, Gly, Thr, Arg, Ala, Cys, Val, Phe, Iso, Leu, and Lys. Besides all, IRPS2A also contained Tyr. Conclusion: This strategy can be used for the isolation and purification of homogeneouspolysaccharides/glycoproteins from IsatidisRadixwhich provides a possible support for the elucidation of the structure and the pharmacologic action of IsatidisRadixpolysaccharides (glycoproteins).

Effect of Heat Treatment on the Amino Acid Profile of Plukenetia conophora Seed Kernel Flours.

The effect of heat treatment on the amino acid composition of Plukenetia conophora (African walnut) seed flours was investigated. The seed kernel of Plukenetia conophora (PC) is eaten roasted or cooked mainly as indigenous snacking nut in Nigeria. The sample was divided into four lots. The 1st was used raw, the 2nd, 3rd and 4th boiled in water (99±1°C) for 45, 90, and 135 min and labeled PCraw, PC45, PC90, and PC135 respectively. The kernels were found to contain amino acids found naturally in plant protein. Glutamic acid (7.88-18.5 g/100 g protein) and aspartic acid (4.86-9.16 g/100 g protein) were the most abundant non-essential amino acids while the essential amino acid ranges were for leucine (4.50-7.80 g/100 g protein), lysine (3.65-7.09 g/100 g protein) and arginine (3.22-6.12 g/100 protein). In addition to leucine other branched-chain amino acids (Isoleucine and valine) were present in high proportion. Cooking progressively decreased the level of all amino acids. For essential amino acid, the percent decrease ranged from 6.07% for lysinein PC45 to 64% for isoleucine in PC135. Protein quality parameters such as: predicted protein efficiency ratio, total amino acid, total essential amino acid, total acidic amino acid, total neutral amino acid, total aromatic amino acid and total sulphur amino acid though decreased by cooking were comparable to those of some plant foods and recommended standards. Based on whole hen’s egg amino acids, FAO amino acid provisional pattern and Food and Nutrition Board/Institute of Medicine (FNB/IOM) pattern for 1-3-year-old child, the percentage adequacy (Essential amino acid score) of most of the essential amino acids in the samplesat all levels of cooking were high. Despite the decreasein the amino acid contents bycooking, the plant food has the potential for giving high quality protein that can be exploited to enhance protein quality in human nutrition and performance in sports.

Production and properties of matallothionein in Cordyceps kyusyuensis under Zn2+ stress / 中草药

Objective: To explore the production and properties of matallothionein (MT) in Cordyceps kyusyuensis under the zinc ion (Zn2+) stress. Methods: By adding ZnSO4 to the culture, the production regularity of MT in C. kyusyuensis under Zn2+ stress was explored. Ultrasonic cell disruption, crude extraction, and gel column chromatography were employed to isolate and purify the MT. The methods of Coomassie brilliant blue, electron spray mass spectrometry (ESMS), Ellman's reagent colorimetry, atomic flame absorption spectrometry, silver saturation method with atomic absorption spectrometry analysis, and amino acid analyzer were used to determine the purity, molecular weight, sulfhydryl content, zinc content, MT content, and amino acid composition, respectively. In addition, the anti-oxidative activity was identified by scavenging capacity of hydroxyl radicals (HO·), DPPH radicals (DPPH·), and superoxide anion radicals (O2·̄). Results: The contents of Zn and MT in mycelia increased strikingly with the increase of Zn2+ concentration (0-15 g/L) in the culture medium. In 10 L fermenter, MT achieved the maximum yield up to 14.335 mg/g mycelium (wet weight) with Zn concentration of 15 g/L in 56 h, and the biomass reached 16.921 g/L. The C. kyusyuensis Zn-MT was obtained with Sephadex G-50 and Sephadex G-25 gel filtration, and dried by vacuum freeze-drying. The molecular weight of the protein was 6 750, one molecule Zn-MT binded six zinc atoms and contained 56 molecules of amino acid, which contained 20 molecules of cysteine and one molecule of histidine. The eliminating action of C. kyusyuensis Zn-MT for HO·, DPPH·, and O2·̄ was better than that of glutathione, and the eliminating action was different in O2·̄ > DPPH· > HO·. The concentration of half clearance rate was 71.49, 80.58, and 100.65 mg/L. Conclusion: According to ultraviolet scan pattern analysis, protein column chromatogram analysis, physicochemical properties, and anti-oxidative activity of C. kyusyuensis Zn-MT, it is similar with the characteristics of standard animal MT, and a certain concentration of zinc sulfate liquid fermentation of C. kyusyuensis could induce the synthesis of Zn-MT, which could provide the reliable theoretical basis for the industrial production.

Prediction of xylanase optimal temperature by support vector regression

Background: Support vector machine (SVM), a novel powerful machine learning technology, was used to develop the non-linear quantitative structure-property relationship (QSPR) model of the G/11 xylanase based on the amino acid composition. The uniform design (UD) method was applied to optimize the running parameters of SVM for the first time. Results: Results showed that the predicted optimum temperature of leave-one-out (LOO) cross-validation fitted the experimental optimum temperature very well, when the running parameter C, ξ, and γ was 50, 0.001 and 1.5, respectively. The average root-mean-square errors (RMSE) of the LOO cross-validation were 9.53ºC, while the RMSE of the back propagation neural network (BPNN), was 11.55ºC. The predictive ability of SVM is a minor improvement over BPNN, but it is superior to the reported method based on stepwise regression. Two experimental examples proved the validation of the model for predicting the optimal temperature of xylanase. Conclusion: The results indicated that UD might be an effective method to optimize the parameters of SVM, which could be used as an alternative powerful modeling tool for QSPR studies of xylanase.

Characteristics of protein from red crab (Chionoecetes japonicus) shell by commercial proteases

This study was performed to examine the characteristics of protein of red crab (Chionoecetes japonicus) shell powder hydrolyzed by commercial proteases. Red crab shell was digested by commercial proteases, such as Protamex (P), Neutrase (N), Flavourzyme (F), Alcalase (A), Protease M (PM) and Protease A (PA). Protein yield analyzed by Biuret assay, absorbance at 280 nm and brix revealed that PA was the enzyme having the highest proteolytic activity. SDS PAGE showed that molecular weight of proteins produced by protease treatments was various and below 150 kDa. Combinational treatment of proteases (PA + P, PA + PM, PA + F, PA + A) was tried whether these increase protein hydrolysis from red crab shell powder compared to a PA single treatment. Soluble protein content was similar, but amino acid concentration by combinational treatments was higher than PA single treatment [PA + P 247.4 mg/g > PA + F (206.4 mg/g) > PA + A (133.4 mg/g) > PA + PM (59.1 mg/g) > PA (54.9 mg/g)]. Amino acid composition by combinational treatments was slightly different. Most abundant essential amino acids were phenylalanine, glycine, alanine, and leucine, whereas tyrosine and cystine were not detected.

Physicochemical properties and biological activity of a recombinant inducible co-stimulator-Ig fusion protein / 第二军医大学学报

Objective: To investigate the physicochemical properties and biological activity of self-prepared fusion protein inducible co-stimulator-Ig. Methods: Acid hydrolysis, edman degradation and peptide mass finger printing were used to determine the amino acid composition, N-terminal 15 amino acid sequences, and peptide mapping. In vivo mixed lymphocyte reaction assay was used for identification of its biological activity. Results: The result of amino acids composition analysis was consistent with the theoretical value of ICOS-Ig. N-terminal 15 amino acid sequences of the product were EINGSANYEMFIFHN, consistent with the theoretical value of ICOS-Ig. Peptide match assay identified six peptides of the product which could match the theoretic maps of ICOS-Ig. ICOS-Ig and CsA noticeably inhibited the proliferation of allo-reactive T cells in vivo. Conclusion: The prepared ICOS-Ig fusion protein has a correct structure and can inhibit the proliferation of allogeneic T cells in vivo, which lays a foundation for quality control of ICOS-Ig fusion protein.

Amino Acid Composition of the α Chain of Hemoglobin and Amino Acid Sequence of it′s Particular Peptide Fragment From 615 Mouse / 生物化学与生物物理进展

The α chain of hemoglobin of 615 mouse was isolated and purified on CM-Celullose-23 colomn chromatography. The N-terminal amino acid of the α chain was valine determined with DABITC/PITC method.The amino acid composition was determined and it was different from the parent(C57BL)in literature on the number of leucine residue,histine residue and valine residue.An undissoluble ‘core’ and dissoluble peptides were found when the α chain of 615 mouse was hydrolysised by trypsin and it was found that the eighth amino acid residue from N-terminal of one particular peptide fragment mutated from valine (C57BL) to leucine.