Analysis on Quality of Sojae Semen Praeparatum Based on Traditional Quality Evaluation / 中国实验方剂学杂志

ObjectiveBased on the quality evaluation experience of "it is better to have a fragrant and strong aroma" summarized by materia medica of past dynasties， the chemical components of Sojae Semen Nigrum（SSN） and Sojae Semen Praeparatum（SSP） were systematically compared and analyzed， and the main fermentation products in different fermentation time were quantitatively analyzed， so as to clarify the transformation law of internal components in the processing process and provide scientific basis for the modern quality control of SSP. MethodUltra performance liquid chromatography-quadrupole tandem time-of-flight mass spectrometry（UPLC-Q-TOF-MS） was used for the structural identification of the chemical constituents of SSN and SSP， and with the aid of Progenesis QI v2.3 software， the negative ion mode was employed for principal component analysis（PCA） pattern recognition， and the data were analyzed with the aid of orthogonal partial least squares-discriminant analysis（OPLS-DA） for two-dimensional data to obtain S-plot， and components with |P|>0.1 were selected as the differential constituents. The contents of isoflavonoids in SSP during fermentation was determined by UPLC， and the samples were taken every 8 h in the pre-fermentation period and every 2 d in the post-fermentation period， and the dynamic changes of isoflavonoid contents in different fermentation stages were analyzed. The contents of amino acids and nucleosides in SSP and SSN from different fermentation stages were quantitatively analyzed by phenyl isothiocyanate（PITC） pre-column derivatization and high performance liquid chromatography（HPLC） gradient elution， and the contribution of flavor substances to the "delicious" taste of SSP was discussed by taste intensity value（TAV）. ResultA total of 19 kinds of differential components were screened out， mainly soybean saponins and isoflavones， and their contents decreased significantly or even disappeared after fermentation. In the pre-fermentation process of SSP， glycoside bond hydrolysis mainly occurred， and isoflavone glycosides in SSN were degraded and converted into the corresponding aglycones， the content of flavor substances such as amino acids increased gradually. In the post-fermentation process， protein degradation mainly occurred， after 8 d of post-fermentation， the content of isoflavones was basically stable， while the total content of amino acids increased by 8-40 times on average. Different amino acids form the special flavor of SSP， such as the TAV of glutamate is always ahead of other flavor substances， and sweet substances such as alanine and valine have made relatively great contributions to SSP. ConclusionBased on the law of constituent transformation， combined with the traditional evaluation index of "fragrant and strong"， it is difficult to control the fermentation degree of SSP by the existing standards in the 2020 edition of Chinese Pharmacopoeia. It is suggested that description of the characteristics of SSP be refined and changed to "fragrant， delicious and slightly sweet"， and at the same time， the post-fermentation index compounds such as glutamic acid， alanine and valine should be added as the quality control indicators of SSP， so as to standardize the production process and improve the quality of SSP.

Use of Multiple Irradiations and Reference Materials as Comparators in Quality Control of Neutron Activation Analysis Data of Biological Samples.

We propose that several different reactor irradiation times followed by assaying of activity for differential counting periods may be employed for quality control (QC) of neutron activation analysis (NAA) data of biological samples. It is also recommended that three to four reference materials (RMs) of similar matrix but from different agencies such as National Institute of Standards and Technology (NIST, USA), International Atomic Energy Agency (IAEA, Vienna), Institute of Nuclear Chemistry and Technology (INCT, Poland), and National Institute of Environmental Studies (NIES, Japan) including a synthetic multielemental primary standard should always be analyzed simultaneously along with the samples. Finally, the mean ± σ values so obtained may be considered as more reliable after statistical analysis. Our analytical data for Na and K in three RMs (SRM 1572, CRM H-9, and Bowen's Kale) using different irradiation periods of 15 m, 2 h, 6 h, and 1 day were comparable with the certified values within error range of + 0.2 to - 2.7%. We report our data for 20 elements in two candidate RMs corn flour (INCT-CF-3) and soya bean flour (INCT-SBF-4) from the INCT, Poland, where Z-score values for most elements are in reasonable range of certified values.

Thorough investigation of non-volatile substances extractible from inner coatings of metallic cans and their occurrence in the canned vegetables.

Since the decline of the use of bisphenol A, the chemistry of the varnishes and coatings which are applied to the inner surfaces of metallic food contact materials is poorly documented. We hypothesised that can coatings are now diverse and bring forth various non-intentionally added substances (NIAS) to be described. Investigating complex components such as NIAS requires demanding non-targeted approaches. We investigated the coatings of 12 vegetable cans from the French market. More than 125 substances were pinpointed, among them 84 oligoester combinations from 8 diols and 4 diacids. Thus, oligoesters were the dominant family. Additives such as epoxidised soybean oil, bisphenol A diglycidyl ether and benzoguanamine derivatives and phenol-formaldehyde oligomers were also identified. A software for exploring databases of theoretical combinations of polyester and phenol-formaldehyde resin components (NIAS-db 1.0) was made available. The stepwise organic synthesis of native and deuterated combinations of neopentyl glycol and isophthalic acid (4 and 8 units, linear and cyclic) enabled a higher confidence level and monitoring in vegetable extracts. Migration of oligoesters averaged 330 µg/kg in the drained vegetables (43-1600 µg/kg). This study sheds light on the need to fulfil a proper risk assessment on this NIAS family (exposure and hazard characterisation).

Effects of a new fermented soya bean meal on growth performance, serum biochemistry profile, intestinal immune status and digestive enzyme activities in piglets.

BACKGROUND: Fermented soya bean meal (FSBM) is believed to have improved nutritional qualities compared with soya bean meal (SBM) and is also cheaper than soya protein concentration (SPC) and fish meal (FM). Therefore, the present study was conducted to compare the effects of FSBM replacing SBM, SPC and FM in diets on growth performance, serum biochemistry profile, short-chain fatty acid concentrations in digesta, intestinal mucosal enzyme activities, intestinal proinflammatory cytokine concentrations and morphology in weaned piglets. One hundred and twenty 28-day-old piglets (Duroc × Landrace × Yorkshire, body weight: 6.73 ± 1.14 kg) were randomly allocated to four treatment diets (six replicate pens with five piglets per pen) containing SBM, SPC, FM or FSBM as the protein source, respectively. RESULTS: Dietary FSBM supplementation improved average daily gain (p < 0.05), gain to feed ratio (p < 0.05), and digestibility of dry matter, gross energy, crude protein and organic matter (p < 0.05) in pigs compared with those fed SBM during 0-14 days and reduced diarrhoea rate (p < 0.05) compared with those fed SBM and FM during 0-14 days. Moreover, pigs fed FBSM had greater IgA and IgM contents and antioxidase activities than those provided SBM and SPC on day 14. In addition, the butyrate concentration in the cecum of pigs fed FSBM was greater than those fed the other diets (p < 0.05), and the trypsin activity in duodenum and jejunum of pigs provided FSBM was greater than those fed SBM (p < 0.05). Moreover, higher villus height (p < 0.05) and villus height to crypt depth ratio (p < 0.05) and lower crypt depth (p < 0.05) in the duodenum of pigs fed FSBM were observed, and pigs fed FSBM had a lower (p < 0.05) TNF-α concentration in jejunum compared with those fed SBM. CONCLUSIONS: In conclusion, dietary FSBM supplementation to replace SBM, SPC and FM could improve piglets' growth performance, intestinal health and immune function.

Influence of an increased content of pea and yellow lupin protein in the diet of pigs on meat quality.

BACKGROUND: The aim of the study was to test the effect of replacing soya beans with pea and yellow lupin seeds in the diet of pigs on meat quality. The meat for the tests was obtained from 60 fattening crossbred pigs F1 (Polish Large White × Polish Landrace) × F1 (Pietrain × Duroc). The animals belonged to three feeding groups depending on the feed used with the total share of soybeans and its reduction. Water holding capacity, colour, and tenderness were measured and visual and tactile evaluation (colour, marbling and firmness) was performed for meat samples collected from the longissimus lumborum muscle. The chemical composition of the meat and the content of minerals were determined. The content of amino acids, fatty acids and cholesterol was determined. RESULTS: There was no significant differences among the assessed physicochemical characteristics of the meat. The obtained meat was of good quality, regardless of the proportion of proteins from legumes in the diet of pigs. The results of the subjective evaluation of meat, its colour, and the content of muscle pigments were uniform in all food groups. Similarly, a uniform, high protein content was found in all tested groups (C-24.98%; E1-24.82%; E2-25.09%) and the content of macro- and micronutrients in the tested meat was equivalent. The profile of fatty acids was not significantly affected by dietary treatment. Palmitoleic acid content was significantly higher (P < 0.05) in the E2 group compared to the E1 group (3.279% compared to 2.844%). The content of amino acids in meat samples was influenced by dietary treatment. Almost all the monitored essential amino acids (threonine, valine, leucine, phenylalanine and lysine) and some of nonessential amino acids' proportion was increased in the experimental groups (E1, E2). CONCLUSION: Replacing soya bean protein in the pigs' diet with legume protein (peas and yellow lupin) did not adversely affect meat quality. This applies to both the physicochemical characteristics, the basic composition and the fatty acid profile. The meat of fattening pigs fed with the highest proportion of pea and lupin in the ration was characterized by more favourable proportion of essential amino acids.

Zebrafish intestinal transcriptome highlights subdued inflammatory responses to dietary soya bean and efficacy of yeast ß-glucan.

Anti-nutritional factors in dietary components can have a negative impact on the intestinal barrier. Here, we present soya bean-induced changes in the intestine of juvenile zebrafish and the effect of yeast ß-glucan through a transcriptomic approach. The inclusion of soya bean meal affected the expression of several intestinal barrier function-related genes like arl4ca, rab25b, rhoub, muc5ac, muc5d, clcn2c and cltb in zebrafish. Several metabolic genes like cyp2x10.2, cyp2aa2, aldh3a2b, crata, elovl4, elovl6, slc51a, gpat2 and ATP-dependent peptidase activity (lonrf, clpxb) were altered in the intestinal tissue. The expression of immune-related genes like nlrc3, nlrp12, gimap8, prdm1 and tph1a, and genes related to cell cycle, DNA damage and DNA repair (e.g. spo11, rad21l1, nabp1b, spata22, tdrd9) were also affected in the soya bean fed group. Furthermore, our study suggests the plausible effect of yeast ß-glucan through the modulation of several genes that regulate immune responses and barrier integrity. Our findings indicate a subdued inflammation in juvenile zebrafish fed soya bean meal and the efficacy of ß-glucan to counter these subtle inflammatory responses.

Protease Inhibitors Protect Bovine Colostrum or Chicken Egg Growth Factors from Pancreatic Enzyme Digestion in AGS Cells or Colitic Rats.

BACKGROUND: Bovine colostrum (BC) and chicken egg contain proteins possessing growth factor activity. Epidermal growth factor (EGF) provides much of the pro-reparative activity within BC. Clinical use of orally administered peptide growth factors is hampered by digestion from pancreatic proteases. OBJECTIVES: We examined whether adding a protease inhibitor [soybean trypsin inhibitor (SBTI) or ovomucoid] protected bioactivity of BC ± egg or EGF alone against pancreatic digestion using in vitro and in vivo models. METHODS: BC, egg, or EGF alone or in combination with trypsin inhibitors were tested for proliferative (Alamar blue) activity using human gastric adenocarcinoma (AGS) cells, prior to and after incubation with HCl/pepsin and trypsin/chymotrypsin. Data were analyzed using 2-factor ANOVA. Eight groups (n = 10) of adult female Sprague-Dawley rats (mean: 188.3 ± 0.8 g) received 20 mg/kg/d of BC + egg, 100 µg/d of EGF, 5 mg/d ovomucoid, or 10.8 mg/d SBTI, alone or in combination (in 1 mL 3% NaHCO3) by gavage for 9 d and dextran sodium sulfate (DSS; 5% in drinking water) for the final 7 d. Histology, microscopic damage score, and myeloperoxidase (MPO) were assessed and analyzed using 1-factor ANOVA. RESULTS: Proliferative activities of BC, egg, or EGF were reduced 40-57% by HCl/pepsin exposure and further reduced 14-24% by chymotrypsin/trypsin. Co-addition of SBTI or ovomucoid truncated the decrease in proliferative bioactivity caused by chymotrypsin/trypsin by 54-100% (P < 0.01). In vivo study showed oral EGF alone or protease inhibitors given alone were ineffective in reducing DSS damage, whereas SBTI with EGF or ovomucoid with BC + egg improved protective effects on weight gain, disease activity score, colonic MPO, and histology damage by 3-4-fold (P < 0.01). CONCLUSIONS: Studies using AGS, cells, and Sprague-Dawley rats showed the protease inhibitors ovomucoid and SBTI protected BC, egg, and EGF against loss of bioactivity due to pancreatic enzymes and, when given with NaHCO3, enhanced colonic protection against DSS damage.

The potential for soybean to diversify the production of plant-based protein in the UK.

Soybean (Glycine max) offers an important source of plant-based protein. Currently much of Europe's soybean is imported, but there are strong economic and agronomic arguments for boosting local production. Soybean is grown in central and eastern Europe but is less favoured in the North due to climate. We conducted field trials across three seasons and two sites in the UK to test the viability of early-maturing soybean varieties and used the data from these trials to calibrate and validate the Rothamsted Landscape Model. Once validated, the model was used to predict the probability soybean would mature and the associated yield for 26 sites across the UK based on weather data under current, near-future (2041-60) and far-future (2081-2100) climate. Two representative concentration pathways, a midrange mitigation scenario (RCP4.5) and a high emission scenario (RCP8.5) were also explored. Our analysis revealed that under current climate early maturing varieties will mature in the south of the UK, but the probability of failure increases with latitude. Of the 26 sites considered, only at one did soybean mature for every realisation. Predicted expected yields ranged between 1.39 t ha-1 and 1.95 t ha-1 across sites. Under climate change these varieties are likely to mature as far north as southern Scotland. With greater levels of CO2, yield is predicted to increase by as much as 0.5 t ha-1 at some sites in the far future, but this is tempered by other effects of climate change meaning that for most sites no meaningful increase in yield is expected. We conclude that soybean is likely to be a viable crop in the UK and for similar climates at similar latitudes in Northern Europe in the future but that for yields to be economically attractive for local markets, varieties must be chosen to align with the growing season.

Enrichment of fermented cassava meal 'mchuchume' with micronutrient ingredients from soya bean flour and Moringa oleifera leaves powder.

BACKGROUND: 'Mchuchume' is a ready to eat fermented cassava food used for breakfast, lunch, and/or snack purposes. Its diverse consumption modes attract many studies, including regarding its fortification with inexpensive food materials rich in beneficial nutritional qualities. This study is aimed at investigating the effect of soya bean flour and Moringa oleifera leaves powder on the micronutrient composition of mchuchume. RESULTS: Enrichment significantly increased iron (0.001 g kg-1 in control to 0.03 g kg-1 in cassava-soya, 0.06 g kg-1 in cassava-moringa and 0.09 g kg-1 in cassava-soya-moringa) and potassium (1.01 g kg-1 in control to 5.14 g kg-1 in cassava-soya, 3.45 g kg-1 in cassava-moringa and 7.43 g kg-1 in cassava-soya-moringa). Compared with the control (0.10 g kg-1 sodium), sodium content either decreased significantly (to 0.09 g kg-1 in cassava-soya) or increased significantly (to 0.37 g k g-1 in cassava-moringa and 0.36 g kg-1 in cassava-soya-moringa). ß-Carotene was below the instrument's limit of detection (<0.2 × 10-6  g mL-1 ) in the control and cassava-soya blends but was detected in cassava-moringa (0.01-0.02 g kg-1 ) and cassava-soya-moringa blends (0.01-0.02 g kg-1 ). CONCLUSION: Mchuchume fortified with both M. oleifera leaves powder and soya bean flour has micronutrients at recommended dietary allowances. The food is recommended for use in management of iron and vitamin A deficiencies, which are endemic not only to inhabitants and refugees of the Kigoma region in Tanzania but also to people in most African countries and other cassava-consuming developing nations. © 2020 Society of Chemical Industry.

Quantification of soya-based feed ingredient entry from ASFV-positive countries to the United States by ocean freight shipping and associated seaports.

African swine fever virus (ASFV) can survive in soya-based products for 30 days with T ½ ranging from 9.6 to 12.9 days in soya bean meals and soya oil cake. As the United States imports soya-based products from several ASFV-positive countries, knowledge of the type and quantity of these specific imports, and their ports of entry (POE), is necessary information to manage risk. Using the data from the International Trade Commission Harmonized Tariff Schedule website in conjunction with pivot tables, we analysed imports across air, land and sea POE of soya-based products from 43 ASFV-positive countries to the United States during 2018 and 2019. In 2018, 104,366 metric tons (MT) of soya-based products, specifically conventional and organic soya bean meal, soya beans, soya oil cake and soya oil were imported from these countries into the United States via seaports only. The two largest suppliers were China (52.7%, 55,034 MT) and the Ukraine (42.9%, 44,775 MT). In 2019, 73,331 MT entered the United States and 54.7% (40,143 MT) came from the Ukraine and 8.4% (6,182 MT) from China. Regarding POE, 80.9%-83.2% of soya-based imports from China entered the United States at the seaports of San Francisco, CA, and Seattle, WA, while 89.4%-100% entered from the Ukraine via the seaports of New Orleans, LA, and Charlotte, NC. Analysis of five-year trends (2015-2019) of the volume of soya imports from China indicated reduction over time (with a noticeably sharp decrease between 2018 and 2019), and seaport utilization was consistent. In contrast, volume remained high for Ukrainian soya imports, and seaport utilization was inconsistent. Overall, this exercise introduced a new approach to collect objective data on an important risk factor, providing researchers, government officials and industry stakeholders a means to objectively identify and quantify potential channels of foreign animal disease entry into the United States.

Monitoring and traceability of genetically modified soya bean event GTS 40-3-2 during soya bean protein concentrate and isolate preparation.

To evaluate DNA fragmentation and GMO quantification during soya bean protein concentrate and isolate preparation, genetically modified soya bean event GTS 40-3-2 (Roundup ReadyTM soya bean, RRS) was blended with conventional soya beans at mass percentages of 0.9%, 2%, 3%, 5% and 10%. Qualitative PCR and real-time PCR were used to monitor the taxon-specific lectin and exogenous cp4 epsps target levels in all of the main products and by-products, which has practical significance for RRS labelling threshold and traceability. Along the preparation chain, the majority of DNA was distributed in main products, and the DNA degradation was noticed. From a holistic perspective, the lectin target degraded more than cp4 epsps target during both of the two soya bean proteins preparations. Therefore, the transgenic contents in the final protein products were higher than the actual mass percentages of RRS in raw materials. Our results are beneficial to the improvement of GMO labelling legislation and the protection of consumer rights.

Lovastatin Production by Aspergillus sclerotiorum Using Agricultural Waste.

RESEARCH BACKGROUND: Lovastatin is a well-known drug used to reduce hypercholesterolaemia. However, the cost of lovastatin production is still high. Therefore, alternative low-cost carbon sources for the production of lovastatin are desirable. EXPERIMENTAL APPROACH: Four different agricultural wastes, namely corn trunks, rice husks, wild sugarcane, and soya bean sludge, were tested separately as substrates to produce lovastatin using a new fungal strain, Aspergillus sclerotiorum PSU-RSPG 178, under both submerged and solid-state fermentation (SSF). RESULTS AND CONCLUSIONS: Of these substrates and cultivation systems, soya bean sludge gave the highest lovastatin yield on dry mass basis of 0.04 mg/g after 14 days of SSF at 25 °C. Therefore, the soya bean sludge was separately supplemented with glucose, wheat flour, trace elements, palm oil, urea and molasses. The addition of the palm oil enhanced the lovastatin yield to 0.99 mg/g. In addition, the optimum conditions, which gave a lovastatin yield of (20±2) mg/g after 18 days of SSF, were soya bean sludge containing 80% moisture (dry basis) at a ratio of soya bean sludge (g) to mycelial agar plugs of 1:4, and a ratio of soya bean sludge (g) to palm oil (mL) of 1:2. Besides, the lovastatin yields obtained from SSF using fresh or dry soya bean sludge were not significantly different. NOVELTY AND SCIENTIFIC CONTRIBUTION: We conclude that A. sclerotiorum PSU-RSPG 178 has a good potential as an alternative strain for producing lovastatin using soya bean sludge supplemented with palm oil as a carbon source.

Experimental dataset on the effect of soaking time and coagulant type on the overall quality of cheese extracted from Ethiopian belessa-95 (Glycine max) soya bean.

Vegetable protein as a replacement for raw materials for the dairy industries is essential to meet the gap between food supply and demand in Ethiopia with particular accentuation of soy protein-based milk and cheese. The experimental dataset presents the extraction of cheese from the soya bean (Belessa-95 or Glycine max). The effects of soaking time and types of coagulants on the overall properties of extracted soy cheese were analyzed. Sensory evaluation was accompanied to highlight the acceptability of the soy cheese product and to further strengthen the significance difference between samples. The experimental method involved pre-treatment as well as extraction process in which four levels of soaking time was taken (12, 24, 36 and 48 h) and three different types of coagulants were used including lemon juice, vinegar, and CaSO4•2H2O. Important properties were tasted to evaluate the best possible amount of soaking time and type of coagulant. These properties were mass yield (%), protein content (%), pH and proximate analysis comprising of moisture content (%), ash content (%) and total solid content (%). The different effects eminent within the values of physical tests are a reflection of the diverse coagulants utilized and different soaking time. Sensory analysis was conducted to further investigate the effect of soaking time and coagulant type. Five semi-trained respondents on a five-point hedonic scale were involved in the process. Data gathered from the sensory evaluation were statistically analyzed using one-way analysis of variance (ANOVA) with a significance level of 5%. Principal component analysis (PCA) was performed on the sensory data to provide additional multivariate graphical presentation.

Identification of loci controlling adaptation in Chinese soya bean landraces via a combination of conventional and bioclimatic GWAS.

Landraces often contain genetic diversity that has been lost in modern cultivars, including alleles that confer enhanced local adaptation. To comprehensively identify loci associated with adaptive traits in soya bean landraces, for example flowering time, a population of 1938 diverse landraces and 97 accessions of the wild progenitor of cultivated soya bean, Glycine soja was genotyped using tGBS® . Based on 99 085 high-quality SNPs, landraces were classified into three sub-populations which exhibit geographical genetic differentiation. Clustering was inferred from STRUCTURE, principal component analyses and neighbour-joining tree analyses. Using phenotypic data collected at two locations separated by 10 degrees of latitude, 17 trait-associated SNPs (TASs) for flowering time were identified, including a stable locus Chr12:5914898 and previously undetected candidate QTL/genes for flowering time in the vicinity of the previously cloned flowering genes, E1 and E2. Using passport data associated with the collection sites of the landraces, 27 SNPs associated with adaptation to three bioclimatic variables (temperature, daylength, and precipitation) were identified. A series of candidate flowering genes were detected within linkage disequilibrium (LD) blocks surrounding 12 bioclimatic TASs. Nine of these TASs exhibit significant differences in flowering time between alleles within one or more of the three individual sub-populations. Signals of selection during domestication and/or subsequent landrace diversification and adaptation were detected at 38 of the 44 flowering and bioclimatic TASs. Hence, this study lays the groundwork to begin breeding for novel environments predicted to arise following global climate change.

Generation of a multiplex mutagenesis population via pooled CRISPR-Cas9 in soya bean.

The output of genetic mutant screenings in soya bean [Glycine max (L.) Merr.] has been limited by its paleopolypoid genome. CRISPR-Cas9 can generate multiplex mutants in crops with complex genomes. Nevertheless, the transformation efficiency of soya bean remains low and, hence, remains the major obstacle in the application of CRISPR-Cas9 as a mutant screening tool. Here, we report a pooled CRISPR-Cas9 platform to generate soya bean multiplex mutagenesis populations. We optimized the key steps in the screening protocol, including vector construction, sgRNA assessment, pooled transformation, sgRNA identification and gene editing verification. We constructed 70 CRISPR-Cas9 vectors to target 102 candidate genes and their paralogs which were subjected to pooled transformation in 16 batches. A population consisting of 407 T0 lines was obtained containing all sgRNAs at an average mutagenesis frequency of 59.2%, including 35.6% lines carrying multiplex mutations. The mutation frequency in the T1 progeny could be increased further despite obtaining a transgenic chimera. In this population, we characterized gmric1/gmric2 double mutants with increased nodule numbers and gmrdn1-1/1-2/1-3 triple mutant lines with decreased nodulation. Our study provides an advanced strategy for the generation of a targeted multiplex mutant population to overcome the gene redundancy problem in soya bean as well as in other major crops.

Multiplex CRISPR/Cas9-mediated metabolic engineering increases soya bean isoflavone content and resistance to soya bean mosaic virus.

Isoflavonoids, which include a variety of secondary metabolites, are derived from the phenylpropanoid pathway and are distributed predominantly in leguminous plants. These compounds play a critical role in plant-environment interactions and are beneficial to human health. Isoflavone synthase (IFS) is a key enzyme in isoflavonoid synthesis and shares a common substrate with flavanone-3-hydroxylase (F3H) and flavone synthase II (FNS II). In this study, CRISPR/Cas9-mediated multiplex gene-editing technology was employed to simultaneously target GmF3H1, GmF3H2 and GmFNSII-1 in soya bean hairy roots and plants. Various mutation types and frequencies were observed in hairy roots. Higher mutation efficiencies were found in the T0 transgenic plants, with a triple gene mutation efficiency of 44.44%, and these results of targeted mutagenesis were stably inherited in the progeny. Metabolomic analysis of T0 triple-mutants leaves revealed significant improvement in isoflavone content. Compared with the wild type, the T3 generation homozygous triple mutants had approximately twice the leaf isoflavone content, and the soya bean mosaic virus (SMV) coat protein content was significantly reduced by one-third after infection with strain SC7, suggesting that increased isoflavone content enhanced the leaf resistance to SMV. The isoflavone content in the seeds of T2 triple mutants was also significantly increased. This study provides not only materials for the improvement of soya bean isoflavone content and resistance to SMV but also a simple system to generate multiplex mutations in soya bean, which may be beneficial for further breeding and metabolic engineering.

Anti-inflammatory effects of functional milk drink enriched with soya bean sprout protein in breastfeeding mothers

@#Introduction: Increased metabolism during pregnancy and breastfeeding results in increased oxidative stress among mothers. However, daily intake of foods containing antioxidants can improve antioxidant and inflammatory status. The objectives of this research were to formulate a soya bean sprout protein milk (SSPM) functional drink; to know its protein and isoflavone contents; and its effect on interleukin 6 (IL-6) level in plasma and breast milk (BM) of breastfeeding mothers. Methods: The study begun with the production of soya bean sprout protein extract (SSPE), followed by preparing five formulated SSPM, namely SSPE, low-calorie sweetener, fructose, skim milk, and salt. The formulated products were subjected to organoleptic test on a scale of 1 (extremely dislike) to 5 (extremely like). Fifty mothers aged 20-35 years who had a newborn up to six months old, in good health condition, and with informed consent were recruited. They were randomly divided into two groups: group I was fed SSPM for two months, 150 ml/day every morning, and group II was treated as placebo. Data were analysed with one-way analysis and paired sample t-test. Results: The preferred SSPM composed of 65.0% SSPE, 1.5% low-calorie sweetener, 6.0% fructose, 27.0% skim milk, 0.5% salt, and containing 13.77±0.001% protein and 229.9±0.001mg/g isoflavone content. Two months after intervention, there was a decrease in the level of IL-6 by 82.5% in the plasma (p=0.015) and 68.1% in BM (p<0.05). Body mass index (BMI) decreased from 22.77 to 20.64 kg/m2 (p=0.019). Conclusion: SSPM is a potential anti-inflammatory agent and has health benefits for breastfeeding mothers.

Proximate composition of fermented cassava meal "mchuchume" fortified with soya bean flour and Moringa oleifera leaves powder.

Mchuchume is a ready to eat meal made from boiled and fermented cassava, which is mainly consumed in Western part of Tanzania. As other fermented cassava foods, it is rich in carbohydrate but deficient of other nutrients. In this study Moringa oleifera leaves powder (MOLP) and soya bean flour (SBF) were used to improve its nutrients. Fortification effects of MOLP and SBF were studied during separate and coetaneous inclusion. Proximate compositions of raw materials and blends were analysed. Mchuchume (control) had moisture, ash, fat, protein, fibre and carbohydrate of 68.138%, 1.090%, 0.589%, 2.068%, 5.738% and 22.367%, respectively, for SBF were 9.522%, 4.990%, 15.436%, 43.807%, 13.751% and 12.486%, respectively and for MOLP were 8.411%, 7.751%, 7.537%, 18.205%, 11.416179% and 46.681%, respectively. Mchuchume-soya had moisture (45.678% and 58.558%), ash (2.398% and 3.664%), fat (2.913% and 5.915%), protein (2.911% and 4.568%), fibre (7.373% and 9.139%) and carbohydrate (22.199% and 27.172%). Mchuchume-moringa had moisture (58.786% and 64.751%), ash (2.248% and 4.184%), fat (0.955% and 1.724%), protein (2.911% and 4.568%), fibre (6.023% and 6.626%) and carbohydrate (29.136% and 30.739%) while mchuchume-soya-moringa had moisture (45.651-58.874%), ash (2.498-4.481%), fat (1.506-4.868), protein (10.722-24.167%), fibre (6.3492-9.408%) and carbohydrate (10.203-21.239%). Mchuchume fortified with both SBF and MOLP are of the most improved nutritive values such that consumption of either of these fortified meals for a day would make an individual to meet his or her recommended dietary allowance for the nutrients analysed at a relatively lesser cost.

Development and Investigation of Lanthanum Sulfadiazine with Calcium Stearate and Epoxidised Soyabean Oil as Complex Thermal Stabilizers for Stabilizing Poly(vinyl chloride).

Lanthanum sulfadiazine (LaSD) was synthesized from sulfadiazine and lanthanum nitrate using water as solvent under alkaline conditions, and was used as a novel rare earth thermal stabilizer to stabilize poly(vinyl chloride) (PVC). The structure of LaSD was characterized by elemental analysis (EA), Fourier transform infrared spectroscopy (FTIR) and thermo- gravimetric analysis (TGA). The influence of lanthanum sulfadiazine with calcium stearate (CaSt2) and epoxidized soybean oil (ESBO) on stabilizing PVC was studied by using the Congo red test, oven discoloration test, UV-vis spectroscopy and thermal decomposition kinetics. The results showed that the addition of LaSD as a thermal stabilizer can significantly improve the initial whiteness and long-term stability of PVC. In addition, the synergies between LaSD, ESBO, and CaSt2 can provide outstanding improvement in the long-term thermal stability of PVC. When the ratio of LaSD/ESBO/CaSt2 is 1.8/0.6/0.6, its thermal stability time is 2193 s which is the best state for stabilizing PVC. Furthermore, comparing the reaction energy (Ea) and the variations in the conjugate double bond concentration in PVC samples, the order of thermal stability of PVC was PVC/LaSD/ESBO/CaSt2 > PVC/LaSD/ESBO > PVC/LaSD. The thermal stability mechanism of LaSD on PVC was studied by the AgCl precipitation method and FTIR spectrum. The results showed that the action of LaSD on PVC was achieved through replacing unstable chlorine atoms and absorbing hydrogen chloride.

Ukrainian Dietary Bread with Selenium-Enriched Soya Malt.

The production of bread with addition of selenium-enriched soya malt was studied. Processing of this soya malt included soaking of the soya beans in the solution of hydroselenite with concentration 1.5 mg Se/L (20 µg of Se per 1 g of soya beans), then 4 days of beans germination at 20 °C, drying at 50 °C until moisture content 8%, separation from the sprouts and grinding. The soya malt was a powder containing 15-18 µg of Se in 1 g. The accumulated selenium was mainly in the protein fraction of soya malt. Addition of selenium-enriched soya malt to leaven intensified activity of yeasts and lactic acid bacteria. The quality of the wheat bread with selenium-enriched soya malt was better than that of the bread in control. The enriched bread had specific pleasant smell and soft texture. The daily intake of 277 g of bread with the selenium-enriched soya malt, which is added in quantity of 1.0-1.75% to mass of plain flour, ensures the consumption of 30-50% of selenium recommended daily allowance for 17 million population of the northern and northwestern Ukraine.