Use of agro-industrial residues for lipase production by Candida viswanathii in a solid-state cultivation system

Lipases are an important group of enzymes that catalyze the hydrolysis of triacylglycerol and there are many industrial applications. The aim of this work was to produce the lipase by the yeast Candida viswanathii using solid state culture with agro-industrial wastes (barley bagasse, corn husk, corncob, soybean seed coat and soybean husk). The biomass pretreatment methods were evaluated, as well as the media supplementation with nitrogen and mixing substrates. Also, the efficiency of olive oil and poultry fat was evaluated on the induction of lipase production, followed by the scale-up from 20 g to 100 g. The enzyme activities in the cultures without pretreatement were higher when soybean seed coat supplemented with both olive oil (7.06 U/gss) and poultry fat (8.40 U/gss) were used. However, the pretreated substrates did not demonstrate a satisfying induction of lipolytic activity. From the nitrogen sources, yeast extract showed an increase of approximately twice the original production with both olive oil (18.12 U/gss) and poultry fat (15.98 U/gss) supplementation. On the scale-up step, the results demonstrated that, for the 20 g culture, the best lipase production was observed on the 7th day (33.52 U/gss), while for the 100 g culture the highest lipase activity was after 5 days (17.88 U/gss). The cultivation of ground soybean skin without pretreatment supplemented with yeast extract as a source of nitrogen, with fresh barley bagasse and poultry fat was the best combination.

Anti-Inflammatory and Anti-Diabetic Effect of Black Soybean Anthocyanins: Data from a Dual Cooperative Cellular System

Obesity is characterized by elevated infiltration of macrophages into adipose tissue, leading to the development of insulin resistance. The black soybean seed coat is a rich source of anthocyanins with antioxidative and anti-inflammatory activities. This study investigated the effects of black soybean anthocyanin extract (BSAn) on obesity-induced oxidative stress, the inflammatory response, and insulin resistance in a coculture system of hypertrophied 3T3-L1 adipocytes and RAW264 macrophages. Coculture of adipocytes with macrophages increased the production of reactive oxygen species and inflammatory mediators and cytokines (NO, MCP-1, PGE2, TNFα, and IL-6) and the release of free fatty acids but reduced anti-inflammatory adiponectin secretion. BSAn treatment (12.5, 25, 50, and 100 μg/mL) alleviated the coculture-induced changes (p < 0.001) and inhibited coculture-induced activation of JNK and ERK signaling (p < 0.01). BSAn also blocked the migration of RAW264.7 macrophages toward 3T3-L1 adipocytes. In addition, treatment with BSAn increased PPARγ expression and glucose uptake in response to insulin in hypertrophied 3T3-L1 adipocyte and RAW264.7 macrophage coculture (p < 0.01). These results demonstrate that BSAn attenuates inflammatory responses and improves adipocyte metabolic function in the coculture of hypertrophied 3T3-L1 adipocytes and RAW264.7 macrophages, suggesting the effectiveness of BSAn for obesity-induced insulin resistance.

Validation of Antiobesity Effects of Black Soybean Seed Coat Powder Suitable as a Food Material: Comparisons with Conventional Yellow Soybean Seed Coat Powder

In this study, we fed obese model mice black soybean seed coat powder (BSCP) and evaluated the antiobesity effects. As a control, normal yellow soybean seed coat powder (YSCP) was used. C57BL/6J, a high-fat diet-induced obesity model mouse, was fed a high-fat diet containing BSCP or YSCP (20% fat) to induce obesity. The results showed that in the BSCP group, it caused significant suppression of body weight gain and suppression of white adipose tissue weight compared with the YSCP group. Moreover, it significantly decreased serum leptin levels, which correlated with visceral fat mass, and increased antidiabetic adipocytokine and adiponectin levels. Therefore, this suggests the pigmented components contained in BSCP have an antiobesity effect in obese model mice. It is suggested that this material, which can be prepared without extraction with an organic solvent and is suitable for use as a food material, could be a functional food material with a practicable antiobesity effect.

Verification of Soybean Seed Coat Colour Specific Markers Reveals I Loci Specific Markers Capable for Distinguishing Genotypes Differing in Seed Coated Colour

Background: In soybean yellow seed coat is preferred in the market, however, colored ones are currently gaining attention because of their medicinal and nutritive values; besides. Hence it is essential to breed varieties with desired seed coat colour. Methods: Twelve genotypes with six each having yellow and black seed coats were screened with fourteen primers linked to seed coat colour governing loci. Result: Out of them twelve primers showed polymorphism. Monomorphism was observed with both T loci specific and two of the three R loci specific primers. However I locus specific primers i.e. SM303, SM305 and TR showed polymorphism shared by their seed coat color. SM303 amplified a 180 bp sized band in yellow seed coated genotypes and a 130 bp band in black seed coated genotypes. SM305 amplified dual bands with a 200bp band being monomorphic and an additional band (192-216 bp range) present in only yellow seed coated genotypes, of which a 208 bp band was shared by four yellow seed coated genotypes. Cold induced seed coat discoloration specific TR primer generated bands of different size ranges in yellow seed coated (336-344 bp) and black seed coated genotypes (300-320), of which a 340 bp band was shared by four yellow seed coated genotypes.

Identification and validation of major QTLs associated with low seed coat deficiency of natto soybean seeds (Glycine max L.)

Key message Two major QTLs associated with low seed coat deficiency of soybean seeds were identified in two biparental populations, and three SNP markers were validated to assist low-SCD natto soybean breeding selection. Abstract Soybean seed coat deficiency (SCD), known as seed coat cracking during soaking in the natto production process, is problematic because split or broken beans clog production lines and increases production costs. Development of natto soybean cultivars with low SCD is crucial to support the growth of the natto industry. Unfortunately, information on the genetic control of SCD in soybean, which is desperately needed to facilitate breeding selection, remains sparse. In this study, two F2 populations derived from V11-0883 × V12-1626 (Pop 1) and V11-0883 × V12-1885 (Pop 2) were developed and genotyped with BARCSoySNP6K Beadchips and F2-derived lines were evaluated for SCD in three consecutive years (2016–2018) in order to identify quantitative trait loci (QTLs) associated with low SCD in soybean. A total of 17 QTLs underlying SCD were identified in two populations. Among these, two major and stable QTLs, qSCD15 on chromosome 15 and qSCD20 on chromosome 20, were detected across multiple years. These QTLs explained up to 30.3% of the phenotypic variation for SCD in Pop 1 and 6.1% in Pop 2 across years. Three SNP markers associated with the qSCD20 were validated in additional four biparental populations. The average selection efficiency of low-SCD soybean was 77% based on two tightly linked markers, Gm20_34626867 and Gm20_34942502, and 64% based on the marker Gm20_35625615. The novel and stable QTLs identified in this study will facilitate elucidation of the genetic mechanism controlling SCD in soybean, and the markers will significantly accelerate breeding for low-SCD soybean through marker-assisted selection.

Black soybean seed coat polyphenols promote nitric oxide production in the aorta through the Akt/eNOS pathway

Background: Black soybean seed coat contains an abundance of flavan-3-ols and possesses various bioregulatory functions. Nitric oxide (NO) is produced by endothelial nitric oxide synthase (eNOS) in vascular endothelial cells and regulates vascular function through vasodilation and the inhibition of platelet aggregation in blood vessels. It has been reported that flavan-3-ols increase NO production, but many previous reports used a high concentration of flavan-3-ols. In the present study, we investigated the effect of flavan-3-ol-rich black soybean seed coat extract (BE) on NO production at a lower concentration that is close to the concentration after permeation through the monolayer of Caco-2 cells.Methods: Human umbilical vein endothelial cells (HUVEC) were incubated with BE, and then NO production in the medium and eNOS phosphorylation in the cells were examined. Intestinal epithelial Caco-2 cells on the upper side of a transwell filter were co-cultured with HUVEC on the basolateral compartment of the transwell apparatus. BE was added from the upper side, and the basolateral medium was collected to measure the concentration of NO and the content of flavan-3-ols. Furthermore, HUVEC were incubated with each flavan-3-ol in order to individuate the most effective compound in BE.Results: BE significantly increased NO production in the medium of HUVEC. When polyphenols in BE were removed from the basolateral medium by ethyl acetate extraction, increased NO production from HUVEC was not observed. Additionally, BE increased phosphorylation of eNOS and Akt in HUVEC. A portion of flavan-3-ols in BE had permeated through intestinal epithelial cells. Among the flavan-3-ols that had permeated, procyanidin C1 had the strongest effect on NO production in HUVEC at the concentration that had permeated the monolayer of Caco-2 cells. Procyanidin C1 (0.05 µM) also induced phosphorylation of eNOS and Akt in HUVEC without affecting the cAMP level. Conclusion: A portion of flavan-3-ols in BE directly promoted NO production through the Akt/eNOS pathway in vascular endothelial cells. These findings suggest that flavan-3-ols in the black soybean seed coat may contribute to improve the vascular function.Keywords: Black soybean seed coat polyphenols; NO; eNOS; Akt; vascular endothelial cells