Virtual screening and directional preparation of xanthine oxidase inhibitory peptides derived from hemp seed protein

The traditional nutritional and medical hemp(Cannabis sativa L.)seed protein were explored for the discovery and directional preparation of new xanthine oxidase inhibitory(XOI)peptides by structure-based virtual screening,compound synthesis,in vitro bioassay and proteolysis.Six subtypes of hemp seed edestin and albumin were in silico hydrolyzed by 29 proteases,and 192 encrypted bioactive peptides were screened out.Six peptides showed to be XOI peptides,of which four(about 67%)were released by elastase hydrolysis.The peptide DDNPRRFY displayed the highest XOI activity(IC50=(2.10±0.06)mg/mL),acting as a mixed inhibitor.The pancreatic elastase directionally prepared XOI hemp seed protein hydrolysates,from which 6 high-abundance XOI peptides encrypted 3 virtually-screened ones including the DDNPRRFY.The novel outstanding hemp seed protein-derived XOI peptides and their virtual screening and directed preparation methods provide a promising and applicable approach to conveniently and efficiently explore food-derived bioactive peptides.

Effects of Hemp seed soft capsule on colonic ion transport in rats

AIM To investigate the effect of Hemp seed soft capsule(HSCC) on colonic ion transport and its related mechanisms in constipation rats.METHODS Sprague-Dawley male rats were randomly divided into three groups: normal group, constipation group and HSSC group. Rats in the constipation and HSSC groups were administrated loperamide 3 mg/kg per day orally for 12 d to induce the constipation model. Then, the HSSC group was given HSSC 0.126 g/kg per day by gavage for 7 d. The normal and constipation groups were treated with distilled water. After the treatment, the fecal wet weight and water content were measured. The basal short-circuit current(Isc) and resistance were measured by an Ussing Chamber. Besides the in vivo drug delivery experiment above, an in vitro drug application experiment was also conducted. The accumulative concentrations of HSSC(0.1 mg/m L, 0.5 mg/m L, 1.0 mg/m L, 2.5 mg/m L, 5.0 mg/m L, 10.0 mg/m L and 25.0 mg/m L) were added to the normal isolatedcolonic mucosa and the Isc was recorded. Further, after the application of either ion(Cl^-or HCO_3^-) substitution, ion channel-related inhibitor(N-phenylanthranilic acid, glybenclamide, 4,4-diisothiocyano-2,2-stilbenedisulfonic acid or bumetanide) or neural pathway inhibitor [tetrodotoxin(TTX), atropine, or hexamethonium], the Isc induced by HSSC was also measured. RESULTS In the constipation group, the fecal wet weight and the water content were decreased in comparison with the normal group(P < 0.01). After the treatment with HSSC, the fecal wet weight and the water content in the HSSC group were increased, compared with the constipation group(P < 0.01). In the constipation group, the basal Isc was decreased and resistance was increased, in comparison with the normal group(P < 0.01). After the treatment with HSSC, the basal Isc was increased(P < 0.05) and resistance was decreased(P < 0.01) in the HSSC group compared with the constipation group. In the in vitro experiment, beginning with the concentration of 1.0 mg/m L, differences in Isc were found between the experimental mucosa(with HSSC added) and control mucosa. The Isc of experimental mucosa was higher than that of control mucosa under the same concentration(1.0 mg/m L, P < 0.05; 2.5-25 mg/m L, P < 0.01). After the Cl^-or HCO_3^-removal and pretreated with different inhibitors(c AMPdependent and Ca^(2+)-dependent Cl^-channels, Na^+-K^+-2 Cl^-cotransporter(NKCC), Na^+-HCO_3^-cotransporter or Cl^-/HCO_3^-exchanger inhibitor), there were differences between experimental mucosa and control mucosa; the Isc of experimental mucosa was lower than that of control mucosa under the same concentration(P < 0.05). Meanwhile, after pretreatment with neural pathway inhibitor(TTX, atropine, or hexamethonium), there were no differences between experimental mucosa and control mucosa under the same concentration(P > 0.05).CONCLUSION HSSC ameliorates constipation by increasing colonic secretion, which is mediated via the coaction of c AMPdependent and Ca^(2+)-dependent Cl^-channels, NKCC, Na^+-HCO_3^-cotransporter or Cl^-/HCO_3^-exchanger.

Emerging natural hemp seed proteins and their functions for nutraceutical applications

With changing dietary habits and increasing awareness of the nutraceutical role of dietary foods,the demand for natural plant proteins and interest in non-traditional protein sources in the food industry are increasing.Industrial hemp,belonging to the plant family Cannabaceae,is cultivated for its fibre and edible seeds.Due to its nutritional value,it has also been used in the food industry and medicine.In particular,hemp seed proteins have drawn considerable attention in both scientific and industrial fields because of their excellent nutraceutical values,superior digestibility,low allergenicity and diverse techno-functional properties.In this review,we provide a summary of the current research progress on the extraction and purification processes,physiochemical properties,nutraceutical functions,and applications of hemp seed proteins.Perspectives in the application of advanced technologies for hemp seed bioactive peptide mining are also discussed.This review provides up-to-date insights into the nutraceutical values,health benefits,and future applications of this emerging plant source protein.

Identification and Expression Analysis of Abscisic Acid Signal Transduction Genes in Hemp Seeds

Abscisic acid(ABA)is involved in regulating diverse biological processes,but its signal transduction genes and roles in hemp seed germination are not well known.Here,the ABA signaling pathway members,PYL,PP2C and SnRK2 gene families,were identified from the hemp reference genome,including 7 CsPYL(pyrab-actin resistance1-like,ABA receptor),8 CsPP2CA(group A protein phosphatase 2c),and 7 CsSnRK2(sucrose nonfermenting1-related protein kinase 2).The content of ABA in hemp seeds in germination stage is lower than that in non-germination stage.Exogenous ABA(1 or 10μM)treatment had a significant regulatory effect on the selected PYL,PP2C,SnRK2 gene families.CsAHG3 and CsHAI1 were most significantly affected by exogenous ABA treatment.Yeast two-hybrid experiments were performed to reveal that CsPYL5,CsSnRK2.2,and CsSnRK2.3 could interact with CsPP2CA7 and demonstrate that this interaction was ABA-independent.Our results indicated that CsPYL5,CsSnRK2.2,CsSnRK2.3 and CsPP2CA7 might involve in the ABA signaling transduction pathway of hemp seeds during the hemp seed germination stages.This study suggested that novel genetic views can be brought into investigation of ABA signaling pathway in hemp seeds and lay the foundation for further exploration of the mechanism of hemp seed germination.

The effect of selected hemp seed protein hydrolysates in modulating vascular function

Reduced bioavailability of nitric oxide along with an increase in inflammatory cytokines,reactive oxygen species,cell adhesion molecules,and platelet hyperactivity are underlying causes of endothelial dysfunction.Bioactive peptides derived from various plants have been shown to attenuate biomarkers of endothelial dysfunction.The objective of this study was to determine the effects of hemp seed protein hydrolysates(HSPH)in modulating biomarkers of endothelial dysfunction using an in vitro cell culture model and an ex vivo platelet activity assay.HUVEC cells were treated with HSPH from commercial variant X and ECO-commercial variant Y,followed by hydrogen peroxide to simulate oxidative stress and inflammation.Biomarkers of endothelial dysfunction were then measured to determine the protective properties of HSPH.Blood samples collected from healthy volunteers were used to determine the effect of the HSPH on ADP and hydrogen peroxide-induced platelet activity,using flow cytometry.The HSPH reduced the production of reactive oxygen species,inflammatory cytokines IL-8,IL-12p70 and IL-1β,and adhesion molecule vascular cell adhesion molecule 1.Additionally,treatment with HSPH also reduced platelet activation marker CD62P ex vivo.The results obtained from this study suggest HSPH alleviates biomarkers of endothelial dysfunction by reducing oxidative stress,inflammation and platelet activation.

Establishment of Tissue Culture Regeneration System in Bama Hemp (Cannabis sativa L.)

[Objectives]This study was conducted to establish a tissue culture regeneration system in Bama hemp(Cannabis sativa L.).[Methods]Using hemp seeds as explants,a regeneration system was established through explant sterilization,callus induction,callus differentiation,and rooting culture.[Results]The results showed that the best sterilization effect was achieved when sterilizing with 75%ethanol for 30 s,followed by 0.1%HgCl 2 solution for 9 min,with a contamination rate as low as 11.4%.In presence of 3 mg/L 2,4-D and 0.1 mg/L6-BA,the callus induction effect from hemp seeds was better.The formula for better differentiation of callus was MS+2.0 mg/L 6-BA+0.2 mg/L NAA.IBA had a promoting effect on the rooting of hemp aseptic plantlets.The highest rooting rate reached 80%when MS+0.3 mg/L IBA were used.[Conclusions]This study established a hemp seed regeneration system to provide technical support for the conservation and breeding of hemp germplasm resources.

火麻籽粕代餐饼干配方及抗氧化活性研究

火麻籽粕是火麻籽提取油脂加工后的副产物,其蛋白质含量为50%左右,油脂含量为5%左右,与火麻籽相比具有高蛋白、低油脂的特性。同时,压榨提取油脂的过程使火麻籽粕具有独特香气,可作为代餐饼干的添加成分。选取不同油厂生产的压榨火麻籽粕1、压榨火麻籽粕2制备代餐饼干,通过单因素实验和正交实验,参考感官评分,确定在100 g低筋面粉中加入6 g压榨火麻籽粕1、20 g黄油、13 g木糖醇、0.2 g碳酸氢钠或8 g压榨火麻籽粕2、25 g黄油、13 g木糖醇、0.2 g碳酸氢钠时,饼干口感、形态最佳。添加了压榨火麻籽粕1、压榨火麻籽粕2的饼干体外消化率均高于80%,与添加了相同比例的大豆粕饼干的体外消化率无显著差异,同时体外消化产物具有1,1-二苯基-2-三硝基苯肼(1,1-diphenyl-2-picrylhydrazyl,DPPH)自由基清除能力。

8个品种大麻籽氨基酸组成分析及蛋白质营养评价

为比较8个不同品种大麻籽蛋白质营养,测定了8个不同品种大麻籽的粗蛋白含量及氨基酸组成、分析呈味氨基酸含量、药用氨基酸含量,采用氨基酸评分(AAS)、化学评分(CS)、氨基酸比值系数分(SRC)、必需氨基酸指数(EAAI)等营养学方法,进行营养价值评价。结果表明,大麻籽中均含有被检测的17种氨基酸,氨基酸种类齐全,其中包括除色氨酸以外的7种必需氨基酸,具有较丰富的营养价值。不同种质大麻籽中氨基酸含量存在差异,氨基酸总量为406.68~616.56 mg/g,必需氨基酸与总氨基酸的比值为0.226~0.234,必需氨基酸与非必需氨基酸的比值为0.291~0.307,都低于联合国粮食及农业组织、世界卫生组织模式参考值0.4和0.6。大麻籽中药用氨基酸占总氨基酸含量的68.87%,G品种中药用氨基酸含量最高,达429.74mg/g,占总氨基酸含量的69.70%。不同种质大麻籽药用氨基酸含量相差较大,但是在总氨基酸中所占比例基本一致。大麻籽样品中谷氨酸、精氨酸、天冬氨酸为含量最高的3种氨基酸,缬氨酸为第一限制性氨基酸。8个品种大麻籽为药用价值非常高的植物蛋白资源。

雌雄配比和种植密度对单雌性工业大麻籽糠产量的影响

以单雌性工业大麻云麻雌1号作为对象,研究不同雌雄配比(雄株比例为10%、20%和30%)种植模式下种植密度(5210、10420、15630株/hm^(2))对麻籽和麻糠产量的影响,进而探讨单雌性工业大麻用于籽糠兼用的生产模式。结果表明,在同一雌雄比例下,种植密度越大,麻籽、麻糠、麻秆及大麻二酚(CBD)理论产量越高;在最大种植密度15630株/hm^(2)下,20%雄株率下的麻籽、麻糠、麻秆及CBD理论产量均高于10%和30%雄株率的产量,其麻籽产量为5738.02 kg/hm^(2),麻糠产量为6087.14 kg/hm^(2),麻秆产量为9396.35 kg/hm^(2),CBD理论产量127.58 kg/hm^(2)。在单雌性工业大麻云麻雌1号籽糠生产实践中,建议配置花期一致的20%左右雄株,保证授粉充足,实现麻籽和麻糠丰产。

GC/ICP-MS法分析不同产地火麻仁中重金属及农药残留

研究黑龙江道地药材火麻仁中的有机氯农药和重金属的残留水平。从黑龙江省内4个片区共采集10批样品,我国南方不同产地采集12批样品,采用电感耦合等离子体质谱法进行重金属的测定,采用气相色谱-质谱法进行农药残留的测定,参照2020版《中国药典》四部通则,对中药材中有机氯农药和重金属的残留进行风险评估。有机氯农药在南方不同产地采集10批样品有所检出;在重金属残留方面,参考《中国药典》对重金属一般限量标准的规定,在南方不同产地采集7批样品超标,其余南方所产火麻仁虽然重金属含量均不超过国标限量标准,但是重金属含量相对于黑龙江省产火麻仁均有较大差异,仍有健康风险。为系统全面精准评价不同产地的火麻仁的质量奠定基础、提供科学依据。

挤压联合酶解制备火麻仁饼粕多肽及其抗氧化性研究

以火麻仁饼粕为原料,经挤压联合酶解制备火麻仁饼粕多肽。以多肽得率为指标,先选择最佳蛋白酶,然后在单因素试验的基础上通过响应面法优化酶解工艺,并研究多肽的抗氧化性。结果表明:最佳蛋白酶为中性蛋白酶;最优酶解条件为酶解时间3.30 h、酶添加量750 U/g(以火麻仁饼粕蛋白干粉质量计)、酶解温度50℃、pH 7.5,在此条件下多肽得率可达13.52%±0.05%。抗氧化性试验表明,挤压联合酶解可有效提高火麻仁饼粕多肽的抗氧化能力。

火麻仁热处理方式对火麻仁乳饮料品质及营养物质的影响

采用热浸(样品W)和烘烤后热浸(样品H)2种方法对火麻仁进行预处理,并制备火麻乳饮料,比较了2种热处理方式对火麻乳饮料的稳定性、感官品质、营养物质和挥发性风味物质的影响。结果显示,样品W的脂肪上浮率(0.84%)、离心沉淀率(9.37%)、可溶性固形物含量(12.57%)和脂质氧化程度(OD532 nm=0.127)较小,颜色偏浅褐色,样品H颜色偏乳白色,感官评分更高。2组样品中的氨基酸含量丰富(2.60 g/100 g左右),样品W中的必需氨基酸占氨基酸总量的42.01%,比样品H高7.13%;两组样品的不饱和脂肪酸相对含量较高(80%左右),其中油酸的相对含量最高(50%以上),样品W的多不饱和脂肪酸含量高于样品H,说明热浸预处理更有利于火麻乳饮料营养物质的保留。样品H和样品W分别检测出25种和21种挥发性风味物质,主要为醛类和醇类化合物,其中含量最高的3种风味物质依次为糠醛、1-辛烯-3-醇和己醛;相比之下,样品H中醇类、杂环类和硫类化合物的相对含量更多。综合来看,2组样品均有较高营养价值,样品W的稳定性更好、必需氨基酸含量和不饱和脂肪酸含量较高,脂肪氧化程度更小,而样品H的风味物质更多,感官评分更高。

栽培措施对张家川工业大麻安全性及籽粒品质的影响

工业大麻是甘肃张家川等地重要的特色作物,但品种退化严重阻碍了该区域工业大麻产业的高质量发展;前期研究对张家川地方品种“张川大麻”进行了提纯复壮,但栽培调控对其安全性及籽粒品质特征的影响还不明确;为此,研究以提纯复壮的“张川大麻”为研究对象,设置种植密度、施肥制度和覆盖方式三因素正交试验,对该品种花叶四氢大麻酚(THC)及大麻二酚(CBD)含量、籽粒蛋白质及油脂含量与产量进行测试;研究发现:提纯复壮显著降低了“张川大麻”的THC含量,且栽培措施对THC含量的影响较小,其生产安全性显著提高;栽培措施对籽粒蛋白质和油脂含量没有显著影响,但施肥方式对单位面积耕地的总产量有显著的影响;为获得较高的投入回报率,建议张家川工业大麻的适宜栽培措施为种植密度45.00×10^(4)plant/hm^(2),配合m(N)∶m(P2O5)∶m(K2O)为70.5∶15∶9。

火麻仁多肽泡腾片的制备工艺优化

通过单因素分析和响应面试验来探究火麻仁多肽泡腾片制备的最优工艺及其质量标准评价.以综合评分为指标,考察辅料的配比与添加量,采用响应面法设计出最优方案.通过筛选最优方案为润滑剂添加量为4.58%、崩解剂添加量为43.35%、崩解剂配比为1.2∶1、矫味剂添加量为1.82%.并对火麻仁多肽泡腾片进行质量标准评价及初步稳定性考察,以多肽泡腾片的外观性状、硬度、崩解时限、pH值、脆碎度、微生物限度等作为检测指标.结果表明,均符合《中国药典》相关规定,具有良好的稳定性.通过单因素和响应面试验对火麻仁多肽进行了系统研究,为火麻仁多肽泡腾片制备提供了理论参考和指导.

不同贮藏条件对火麻种子生理生化及其活力的影响

本研究探究了不同含水量和贮藏温度处理下,火麻种子生理指标的变化情况,为火麻种质资源保存方面提供科学依据。以云南楚雄和黑龙江大庆的火麻种子为材料,分别设置6个不同含水量贮藏于7℃、-15℃、-30℃、-80℃和常温25℃左右条件下6个月,分析火麻种子过氧化氢酶(CAT)、过氧化物酶(POD)、脱氢酶(DHA)、丙二醛(MDA)、相对电导率、萌发率的变化情况。结果表明,不同含水量和不同贮藏温度对火麻种子的生理指标有影响,云南楚雄和黑龙江大庆的火麻种子均随种子含水量的增加其DHA活性呈先上升后降低趋势。大庆种子随种子含水量的增加其CAT活性呈上升趋势。楚雄种子和大庆种子MDA含量均随含水量增加有上升趋势。两地种子均随含水量的增加,种子的相对电导率增加。此外,与其他贮藏温度相比,楚雄种子在7℃下各含水量均维持较高萌发率。本研究结果表明,贮藏温度为7℃,楚雄种子含水量在5.13%~6.16%左右,大庆种子含水量在4.03%~5.26%左右时火麻种子活力较好,能提高种子品质,在火麻种子贮藏条件选择方面具有重要的参考价值。

纤维素酶辅助碱提汉麻籽分离蛋白工艺优化

汉麻籽分离蛋白(Hemp Seed Protein Isolate,HPI)是汉麻籽深加工过程中的产物之一,尚未被完全利用开发。本文采用酶法水解脱脂汉麻籽粉辅助碱提HPI,以提取率和沉淀率为指标,设计单因素实验和正交实验,研究碱提料液比、碱提温度、碱提pH值、酶解pH值对纤维素酶辅助碱提酸沉法提取汉麻籽蛋白法的影响,并评估对比产品质量。结果表明,最佳工艺条件为酶解pH=4.0,料液比1∶20,碱提温度45℃,碱提pH=10.0。此条件下制备HPI沉淀率达到了77.9%,蛋白含量为83.69%。

火麻仁蛋白的反胶束法提取工艺、理化性质及消化率研究

[目的]利用反胶束法提取火麻仁蛋白,进行工艺优化并分析所得蛋白的理化性质。[方法]对前萃取液2-乙基己基琥珀酸酯磺酸钠(AOT)质量浓度、加水量、缓冲液pH值、KCl浓度、料液比、前萃取温度、前萃取时间与后萃取液pH值、KCl浓度、后萃取时间、后萃取温度等影响蛋白提取率的关键因素进行优化,考察所得蛋白的溶解性、持水性、持油性与消化特性。[结果]反胶束最佳前萃取条件为AOT质量浓度0.0904 g/mL、缓冲液pH值6.54、加水量0.1058 mL/mL、KCl浓度0.05 mol/L、料液比1∶30(g/mL)、前萃取时间60 min、前萃取温度50℃,前萃取率达71.38%;最佳后萃取条件为pH值7.5、KCl浓度1.5 mol/L、后萃取时间45 min、后萃取温度27℃,后萃取率达80.59%,总萃取率为57.53%。经透析、冷冻干燥所得蛋白为白色、蓬松粉末,纯度达98.12%,其溶解性随pH值变化显著,持水性为0.71 g/g,持油性为3.57 g/g,消化率为93.83%。[结论]火麻仁蛋白可用反胶束法提取,较好的持油性、高消化率等特性使火麻仁蛋白在食品应用中具有独特的优势。

火麻仁对小麦粉面团的流变学特性和面包品质的影响

本文以火麻仁粉、小麦粉为主要原料制备面包,通过粉质仪、拉伸仪探讨火麻仁粉添加量对混合粉面团流变学特性的影响,并依据质构分析和感官评价判断火麻仁小麦面包的品质。结果表明,添加火麻仁粉会对面团的稳定时间产生负面影响,面团弱化度、吸水率和粉质质量指数都有不同程度的降低;添加8%火麻仁粉的面团醒发45 min、90 min时的延展性和持气能力都优于对照组,且不同醒发时间组的拉伸能量均达到最大。火麻仁粉添加量为8%时的面包蓬松饱满,气孔均匀,面包芯柔软且富有弹性,浓郁的小麦香气中含有火麻仁特有的清香。相关结果可为火麻仁在面制品中的应用提供思路。

打顶和喷施乙烯利对工业大麻籽粒和麻糠产量的影响

为探究提高雌雄异株工业大麻品种籽粒和麻糠产量的农艺措施,以云麻8号为试验材料,开展大田试验研究开花前打顶和喷施乙烯利处理(0、100、200、400 mg/L)对籽粒和麻糠产量的影响。结果表明,打顶处理会导致雌株的籽粒和麻糠产量下降。喷施乙烯利显著增加了雌株数量,雌雄株数比值由1.04提高到1.60。随喷施乙烯利浓度的增加,雌株的籽粒和麻糠产量均先升高后降低。喷施200 mg/L乙烯利溶液的雌株籽粒和麻糠产量最高,分别为2158.4 kg/hm^(2)和1978.4 kg/hm^(2),较0 mg/L处理分别增加了50.8%和28.5%,较400 mg/L处理分别高283.5 kg/hm^(2)和549.0 kg/hm^(2)。喷施乙烯利还会导致部分雄株分化出雌花并结实,喷施400 mg/L乙烯利时雄株籽粒产量最高,达到910.5 kg/hm^(2),但雄株麻糠产量在喷施200 mg/L乙烯利时最高,为427.5 kg/hm^(2)。综合雌株和雄株的产量,喷施400 mg/L乙烯利的籽粒产量最高,喷施200 mg/L乙烯利的麻糠产量最高。

巴马火麻油对大鼠抗氧化功能的影响

为研究巴马火麻油对大鼠抗氧化功能的影响,依据《关于印发抗氧化功能评价方法等9个保健功能评价方法的通知》(国食药监保化[2012]107号)附件1抗氧化功能评价方法,采用巴马火麻油对成年大鼠进行实验,设溶媒对照组、空白对照组及火麻油高、中、低3个剂量组,连续灌胃30 d。结果表明,受试样品组肝组织匀浆和血清中的丙二醛低于溶媒对照组,差异显著(P<0.05),肝组织匀浆中的超氧化物歧化酶活性极显著高于溶媒对照组(P<0.01);受试样品组肝组织匀浆和血清中的还原型谷胱甘肽含量高于溶媒对照组,差异极显著(P<0.01)。这表明在本实验条件下,动物实验的脂质过氧化产物、抗氧化酶、抗氧化物质3项指标阳性,表明火麻油具有抗氧化功能。