薏苡仁多菌发酵液的菌种优选及其抑制黑色素生成的作用研究

以薏苡仁作为发酵基质,确定利于提高发酵液体外活性的较优乳酸菌种,并分析优势乳酸菌种薏苡仁发酵液对斑马鱼胚体黑色素生成的抑制作用。通过比较分析乳酸乳球菌（Lactococcus lactis）、嗜热链球菌（Streptococcus thermophilus）和保加利亚乳杆菌（Lactobacillus bulgaricus）3种单一乳酸菌和三者复合乳酸菌的薏苡仁发酵液的还原糖、总酚、游离氨基酸、蛋白、总酸和乳酸含量等理化指标及体外羟自由基清除能力和酪氨酸酶活抑制率确定较优发酵菌种,采用高通量测序测定发酵过程中微生物菌群结构;利用斑马鱼模型研究发酵液对黑色素生成的抑制作用。研究结果表明,采用乳酸乳球菌、嗜热链球菌和保加利亚乳杆菌3种乳酸菌复合发酵比单一乳酸菌发酵更具优势。使用以上菌种复合发酵薏苡仁过程中,乳酸乳球菌和嗜热链球菌为发酵前期优势菌群,发酵中后期则以保加利亚乳杆菌为优势菌群。经复合乳酸菌发酵后,薏苡仁发酵液的羟自由基清除率和酪氨酸酶活抑制率分别提高了20.82%和87.26%;斑马鱼模型实验结果表明,薏苡仁发酵液可以显著减少斑马鱼体表黑色素分布,当使用含量为2.0%时,对黑色素...     

两株真菌降解菜籽饼中植酸的研究

利用选择性培养基从土壤中分离到两株能降解植酸的丝状真菌。这些菌株能利用肌醇作为唯一的碳源和能源而生长。在液态发酵中植酸的降解率分别为74．4％和95．0％;在固态发酵中植酸的降解率为40％左右。某些金属离子对菌株的降解率的提高具有一定的促进作用。对温度、pH和水分等影响因子也进行了初步的探讨。经初步鉴定,这两株菌株中有一株为拟青霉（Paecilomycessp）,另一株为青霉（Penicilliumsp．）,它们均不产黄曲霉素素。     

冬虫夏草提取物的美白作用

以冬虫夏草提取物(Chinese cordyceps extract)为研究对象,通过蘑菇酪氨酸酶活性抑制试验和小鼠皮肤黑色素瘤细胞(B16-F10)黑素合成抑制试验考察冬虫夏草提取物的美白活性。结果显示冬虫夏草提取物(质量浓度40～200 mg/m L)呈剂量依赖性抑制蘑菇酪氨酸酶的活性,且在安全剂量(质量浓度0.1～0.5 mg/m L)下显著抑制小鼠皮肤黑色素瘤细胞(B16-F10)内的黑色素合成(P0.05)。说明冬虫夏草提取物能通过抑制酪氨酸酶活性有效阻滞黑色素的合成,从而实现美白作用。     

黑色素形成机理、生物学功能和应用开发的研究进展

黑色素（melanin）是一类化学结构极其复杂、非均质的酚类或吲哚类物质聚合体,是自然界中最为丰富的天然色素,广泛存在于各种动物、植物和微生物中。根据合成途径和中间代谢产物的不同,黑色素主要可分为为真黑素（eumelanin）、棕黑素（pheomelanin）、异黑色素（allomelanin）三大类。其中异黑色素又包括脓黑素（pyomelanin）、1,8-二羟基萘（dihydroxynaphalene, DHN）黑色素等。基于黑色素的生化功能,它们在工业、医药、农业中都有广泛用途,是重要的生物资源。本文主要介绍天然黑色素在动植物和微生物中的合成途径、生物学功能以及有潜力的获取方法和应用前景。     

基于全基因组数据分析香菇黑色素合成途径及相关基因

香菇菌丝转色是决定栽培香菇产量和品质的关键因素之一。弄清菌丝转色过程中黑色素合成途径和关键基因对于解析香菇菌丝转色机理具有重要意义。本研究利用一组可亲和配对的香菇原生质体单核菌株SP3、SP30全基因组数据进行同源比对,对香菇黑色素合成途径进行注释并对黑色素合成关键基因进行挖掘。结果显示,香菇基因组中缺少庚烯酮水解酶基因、小柱孢酮脱水酶基因和对羟苯丙酮酸双加氧酶基因,推测香菇菌株不存在典型的DHN-黑色素和脓黑色素合成途径,但可以合成DOPA黑色素、GHB黑色素、PAP黑色素和儿茶酚黑色素。同时进一步对香菇黑色素合成中限速酶——酪氨酸酶进行生物信息学分析,研究表明香菇菌株SP3、SP30中酪氨酸酶基因编码蛋白为稳定的亲水蛋白,含有2个与酪氨酸酶催化活性相关的铜离子结构域,不含有分泌型信号肽,不含跨膜结构域且定位在细胞质。该研究结果为香菇黑色素合成遗传基础、菌丝转色机理及影响因素研究提供了重要参考。     

金线莲抑制斑马鱼黑色素形成的活性组分筛选及机理研究

为探索金线莲中对黑色素形成具有抑制效果的活性组分,本研究对金线莲进行分离、提取,获得总提组、醇沉组与醇提组,利用斑马鱼筛选金线莲具有美白作用的活性组分。将受精后0.75 h的斑马鱼胚胎分别暴露于不同浓度的金线莲总提组、醇沉组、醇提组,72 h时观察结果表明,金线莲醇提物能有效抑制斑马鱼胚胎黑色素和黄色素沉着,浓度越高抑制效果越明显,且不影响胚胎生长发育。进一步采用半定量PCR和整胚原位杂交技术定量和定性地检测黑色素形成相关基因mRNA表达,结果表明金线莲醇提物可以有效降低silv、tyr和tyrp1a等黑色素合成相关基因的转录水平,且具有浓度依赖关系。通过检测酪氨酸酶活性显示,加入醇提物的实验组其酪氨酸酶活性随着处理浓度升高而逐渐降低。此外,在黑色素已经大量形成的情况下,金线莲醇提物仍可通过下调黑色素合成相关基因的mRNA表达及酪氨酸酶活性来抑制黑色素的形成,并且这种抑制效果可在金线莲醇提物撤除后得到恢复。上述实验结果表明,金线莲醇提物能显著抑制斑马鱼黑色素的形成,本文为金线莲在美白产品领域的开发和应用方面提供了有力支撑。     

miR-411a-3p抑制羊驼黑色素细胞黑色素的生成

microRNA-411a-3p（miR-411a-3p）在不同毛色羊驼皮肤中差异表达,提示其可能在毛色形成中起调控作用。为证明miR-411a-3p在羊驼皮毛黑色素形成中的作用,本研究通过生物信息学预测及靶基因搜索,发现胰岛素样生长因子1受体（IGF1R）是miR-411a-3p的靶基因,继而构建了含Igf1r mRNA 3′-UTR的荧光素酶报告基因载体。报告基因转染结合报告酶活性测定证明,Igf1r是miR-411a-3p的靶基因。原位杂交显示,miR-411a-3p主要在羊驼黑色素细胞胞质中表达,提示其可能在黑色素细胞中具有重要作用。qRT-PCR揭示,棕色和黑色羊驼皮肤中miR-411a-3p表达量明显低于白色羊驼。qRT-PCR联合蛋白质印迹分析显示,在羊驼黑色素细胞过表达miR-411a-3p,伴随Igf1r下调,小眼畸形相关转录因子（Mitf）依赖的毛色基因酪氨酸酶（Tyr）、酪氨酸酶相关蛋白-2（Tyrp2）及黑色素表达水平明显下调。上述结果提示,miR-411a-3p可通过靶向抑制Igf1r负调控羊驼黑色素黑色素合成。该结果将加深我们对羊驼毛色形成机制的认识。     

Influence of Rapeseed Cake on Heavy Metal Uptake by a Subsequent Rice Crop After Phytoextraction Using Sedum plumbizincicola

A glasshouse pot experiment was conducted to study the effects of phytoextraction by Sedum plumbizincicola and application of rapeseed cake (RSC) on heavy metal accumulation by a subsequent rice (Oryza sativa L.) crop in a contaminated paddy soil collected from east China. After phytoextraction by S. plumbizincicola the soil and brown rice Cd concentrations effectively declined. After phytoextraction, RSC application reduced brown rice Cd concentrations in the subsequent rice crop to 0.23–0.28 mg kg^?1, almost down to the standard limit (0.2 mg kg^?1). After phytoextraction and then application of RSC, the soil solution pH, dissolved organic carbon (DOC) and dissolved organic nitrogen (DON) concentrations increased during early stages of rice growth resulting directly and indirectly in lowering the bioavailability of the heavy metals. Thus the grain yield of the subsequent rice crop increased and the heavy metals in the brown rice declined significantly. In this contaminated acid soil, growing the hyperaccumulator S. plumbizincicola and rice in rotation together with RSC application may therefore be regarded as a viable strategy for safe grain production and bioremediation.     

红景天提取物提高高强度跑台运动小鼠的抗氧化能力

为了考察红景天提取物对高强度跑台运动小鼠的抗氧化能力的影响,本研究将30只昆明小鼠随机分成对照组、模型组和红景天提取物组,每组10只。红景天提取物组小鼠按照500 mg/kg bw的剂量灌胃红景天提取液(2 m L)。对照组和模型组小鼠灌胃等体积的蒸馏水,共灌胃4周。采用硫酸蒽酮比色法检测小鼠肝脏和肌肉组织糖原的含量;采用硫代巴比妥酸比色法检测小鼠骨骼肌组织中的丙二醛(MDA)水平;采用RT-PCR和Western blotting检测骨骼肌组织中的超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)和过氧化氢酶(CAT)的表达水平;采用苏木精和伊红(HE)染色评价骨骼肌病理改变。研究显示,模型组小鼠的跑台运动时间显著低于红景天提取物组(61.32 min vs 83.22 min,p<0.05);与模型组相比,红景天提取物组小鼠骨骼肌的炎性细胞浸润明显减轻,肌纤维排列明显改善;红景天提取物组的肝糖原和肌糖原含量均显著高于模型组;红景天提取物组小鼠骨骼肌组织中的MDA水平显著低于模型组;红景天提取物组的SOD、GSH-Px和CAT m RNA和蛋白表达水平均显著高于模型组;红景天提取物可通过上调抗氧化酶表达来增加抗氧化能力,减弱骨骼肌损伤,并增加机体的抗疲劳能力。     

Anti-melanogenesis potential of a new series of Morita-Baylis-Hillman adducts in B16F10 melanoma cell line

Melanin is a natural polymer pigment which provides skin photoprotection against ultraviolet radiation. An excessive synthesis of melanin leads to hyperpigmentation disorders. Tyrosinase catalyzes the rate limiting steps on melanogenesis. Therefore, tyrosinase inhibitors have potential applications in medicine and cosmetic fields. We carried out herein the screening of a family of cyclic Morita-Baylis-Hillman adducts (MBH) to find out their effects on tyrosinase activity and on melanogenesis in murine melanoma B16F10 cell line. Kinetic analysis of tyrosinase inhibition showed that compounds 1a (2-hydroxymethyl) cyclohex-2-enone) and 3f (diethyl (1-(6-oxocyclohex-1-en-1-yl) ethyl-phosphonate) were competitive inhibitors, whereas the compound 2b (6-oxocyclohex-1-en-1-yl) ethyl acetate) was a non-competitive one. Additionally we have found that (1a, 2b and 3f) compounds had a strong melanogenesis inhibition effect in isobutylmethylxanthine (IBMX)-treated murine melanoma B16F10 cells when tested at low and non cytotoxic dose (10–50 µM), by attenuating the melanin production, intracellular tyrosinase activity and tyrosinase expression. Thus, we suggest that these compounds could be used as effective skin-whitening agents.     

菜籽降压肽双酶水解液脱色工艺的研究

用活性炭对菜籽降压肽双酶水解液进行脱色处理,采用单因素试验以及单因素试验基础上的正交试验法。考察活性炭用量、pH、温度、时间对水解液脱色率、肽损失率、活性及ACE抑制率的影响。正交试验优化结果表明,菜籽降压肽双酶水解液脱色最佳工艺为活性炭的用量为1%、脱色pH值为3.0、脱色温度为80℃、脱色时间为50 min,在此条件下脱色率为85.52%、ACE抑制率为57.83%、活性损失率为5.69%。     

Determination of Apparent Ileal Amino Acid Digestibility in Rapeseed Meal and Cake Processed at Different Temperatures Using the Direct and Difference Method with Growing Pigs

Studies were conducted with ten barrows, average initial body weight 34.5 ± 2.1kg, fitted with a T-cannula at the distal ileum, to study the accuracy of determination of the apparent ileal digestibility (AID) values of crude protein (CP) and amino acids (AA) in rapeseed meal and cake and the effects of processing, using the difference method. Five corn starch-based diets in the studies were formulated to contain 17.7% CP and based on soybean meal, prepress-extraction rapeseed meal, prepress-extraction rapeseed meal plus soybean meal, high-temperature press rapeseed cake plus soybean meal, or low-temperature press rapeseed cake plus soybean meal as the sole source of dietary protein. The design was an incomplete Latin Square involving two three-week periods and five-treatments. It was found that the AID values of CP and most AA determined with the difference or direct method were significantly lower in rapeseed meal or cakes than soybean meal. The AID values of CP and most AA in prepress-extraction rapeseed meal, high-temperature press or low-temperature cakes determined with the difference method were no difference from those in prepress-extraction rapeseed meal determined with the direct method. The AID values of CP and AA in rapeseed meal and cake determined with the difference method were accurate, when the contribution of CP and AA from rapeseed was more than 50%. The AID values of CP and AA (especially lysine) were lower in the high-temperature press rapeseed cake than in the low-temperature press cake or the prepress-extraction meal.     

蚕豆花水提取物的UPLC-ESI-QTOF质谱联用分析及抗氧化活性研究

为阐明蚕豆(Vicia faba L.)花水提取物的药效作用基础,采用超高效液相色谱-高分辨飞行时间质谱(UPLC-ESI-QTOF-MS)联用技术对水提取物进行分析鉴定,并测定其抗氧化活性。结果表明,从蚕豆花水提取物中鉴定出178种化学成分,其中相对含量大于0.01%的成分有107种,相对含量较高的成分有野黄芩素-7-新橘皮糖苷、3-吲哚基-β-d-葡糖苷酸环己胺盐、螺环内脂E、芦丁、海藻酸丙二醇酯、山柰酚-3-α-d-半乳糖甙、4,8-二羟基喹啉-2-甲酸等。主要成分以黄酮类和生物碱类为主,分别有16种和17种,相对含量分别为36.53%和34.33%。清除DPPH自由基的IC50值为26.5 μg/mL,清除ABTS+自由基的IC50值为25.0 μg/mL。黄酮类和生物碱类是蚕豆花发挥功效的主要物质基础,蚕豆花水提取物具有较强的抗氧化活性,蚕豆花具有较高的应用价值。     

优质油菜籽粕蛋白质分类研究

油菜是我国大面积种植的油料作物 ,油菜籽富含油脂和蛋白质。其含硫氨基酸含量高于大豆蛋白 ,必需氨基酸含量平衡[1 ] ,因此菜籽蛋白有良好的营养特性。但常规油菜籽饼粕含较高的硫代葡萄糖甙和酚类物质 ,长期以来籽粕蛋白没有被利用作饲料或食品添加剂。我们用双低 [2 ] (低芥酸、低硫甙 )优质油菜饼粕为材料 ,根据其蛋白在不同溶剂的溶解性分离 [3] ,测定了不同组分蛋白及其含量 ,并对各组分蛋白氨基酸含量也进行了测定 ,为说明优质油菜饼粕蛋白富含各种必需氨基酸和其良好的开发前景提供了依据。1 　实验材料及方法1 .1 　材料中双 1 1 9…     

五倍子提取物抑菌效果及稳定性研究

采用琼脂打孔法和平板涂布法,观察五倍子提取物对常见致病菌的抑菌效果,并以金黄色葡萄球菌、大肠杆菌的抑菌环直径为参考指标,考察紫外光照时间、温度及介质pH值对五倍子提取物抑菌活性的影响。结果表明,五倍子提取物对革兰氏阳性菌的抑菌效果明显强于革兰氏阴性菌,在稳定性研究过程中发现,光照时间、温度及介质pH值对五倍子提取物的抑菌效果影响较大,因此在选用五倍子提取物作植物抑菌剂时应现配现用。     

Lightening Effect on Ultraviolet‐Induced Pigmentation of Guinea Pig Skin by Oral Administration of a Proanthocyanidin‐Rich Extract from Grape Seeds

Antioxidants such as vitamins C and E have been reported to inhibit the progression of ultraviolet (UV) radiation‐induced pigmentation in the skin of hairless mice. However, little is known of the lightening effect of proanthocyanidin, a powerful polyphenolic antioxidant, on UV‐induced pigmentation of the skin. We investigated the lightening effect of oral administration of a proanthocyanidin‐rich grape seed extract (GSE) using guinea pigs with UV‐induced pigmentation. These pigmented guinea pigs were fed diets containing 1% GSE or 1% vitamin C (w/w) for 8 weeks. GSE‐feeding had an apparent lightening effect on the guinea pigs’ pigmented skin. Histologic evaluation demonstrated a decrease in the number of 3,4‐dihydroxyphenylalanine (DOPA)‐positive melanocytes as well as 8‐hydroxy‐2′‐deoxyguanosine (8‐OHdG)‐positive, Ki‐67‐positive, proliferating cell nuclear antigen (PCNA)‐positive melanin‐containing cells in the basal epidermal layer of the UV‐irradiated skin in GSE‐fed guinea pigs. In contrast, these parameters did not change in the skin of vitamin C‐fed or control guinea pigs. GSE inhibited the activity of mushroom tyrosinase and also inhibited melanogenesis without inhibiting the growth of cultured B16 mouse melanoma cells. In conclusion, we demonstrated that oral administration of GSE is effective in lightening the UV‐induced pigmentation of guinea pig skin. This effect may be related to the inhibition of melanin synthesis by tyrosinase in melanocytes and the reactive oxygen species (ROS)‐related proliferation of melanocytes.     

菜籽饼肥对香蕉枯萎病的防效及其对土壤细菌的影响

研究菜籽饼肥对香蕉枯萎病的防治效果,探究菜籽饼肥施用后对土壤细菌群落的影响.设置2组实验,CN组为菜籽饼肥处理组,YN组为对照实验组.种植120d后,统计各组的发病率;利用qPCR技术检测2组中的FOC4孢子数量;采集2组土壤,利用Illumina Miseq高通量测序平台对CN处理组与YN对照组的土壤细菌群落结构和差异进行分析对比.实验研究发现:(1)施菜籽饼肥的CN处理组枯萎病发病率比YN对照组低63.33％;施用菜籽饼肥的处理土壤中FOC4的数量由2.94×104个/克土下降为1.96×103个/克土,YN对照组土壤中FOC4孢子的数量由2.94×104个/克土上升为4.55×105个/克土,CN处理组较YN对照组枯萎病菌数量下降了 2个数量级;(2)土壤细菌宏基因组微生物分类测序结果显示,CN处理组的Alpha多样性指数均优于YN对照组,这表明施用菜籽饼肥后增加了土壤细菌群落的丰度及多样性;(3)在门水平上,菜籽饼肥处理增加了变形菌门(Proteobacteria)和拟杆菌门(Bacteroidetes)的相对丰度,同时也降低了酸杆菌门(Acidobact-eria)、厚壁菌门(Firmicutes)和Patescibacteria等菌门的相对丰度;在属水平,CN处理组中的不动杆菌属(Aci-netobacter)、水杆菌属(Aquabacterium)、热单胞菌属(Thermomonas)、产黄杆菌属(Rhodanobacter)、假单胞菌属(Pseudomonas)和奥托氏菌属(Ottowia)等菌属的相对丰度较YN对照组有明显提高,而酸热菌属(Acidother-mus)、Bryobacter菌属以及Acidipila等菌属相对丰度较YN对照组有所减少.施用菜籽饼肥可以改变土壤中细菌的群落结构,显著降低土壤中FOC4孢子数量,从而降低香蕉枯萎病的发病率.本研究结果为菜籽饼肥用于香蕉枯萎病综合防控提供了理论依据.