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1.
为确定维生素C二步发酵中巨大芽孢杆菌(伴生菌)芽孢形成对氧化葡萄酸杆菌(产酸菌)产酸的影响,本研究通过对巨大芽孢杆菌生长特性分析,选取培养12h(未形成芽孢)和36h(芽孢大量形成)巨大芽孢杆菌B.m2980,检测其胞外液、胞内液以及混合液对产酸菌生成2-酮基-L-古龙酸的影响。结果表明,在未开始形成芽孢时,伴生菌胞外液、胞内液及混合液对产酸菌的生长和产酸有较低的促进作用,其中胞内液的促进能力大于胞外液;在芽孢生成后,胞外液以及混合液对产酸菌生长和产酸的促进能力显著提高。  相似文献   

2.
【目的】在酿酒酵母中异源表达双孢蘑菇来源的酪氨酸酶基因PPO2,并研究酪氨酸酶在酿酒酵母胞内及胞外的酶学特性。【方法】提取双孢蘑菇总RNA,通过RT-PCR克隆酪氨酸酶基因PPO2,构建表达载体pSP-G1-PPO2,并转化至酿酒酵母进行表达,采用镍亲和层析纯化蛋白并研究其酶学性质。【结果】在酿酒酵母中正确表达了大小为65 kDa的酪氨酸酶蛋白。重组酶能催化底物酪氨酸产生黑色素。体外活性测定表明,酪氨酸酶催化最适温度为45°C,以酪氨酸和多巴为底物时最适pH分别为7.0和8.0。在酿酒酵母中测得底物酪氨酸浓度低于2.5 mg/mL时,黑色素的产量与底物浓度呈现正相关性。【结论】来源于双孢蘑菇的酪氨酸酶基因PPO2在酿酒酵母中成功表达,重组酶具有良好的酶学特性。利用酪氨酸酶产物黑色素的产量与底物浓度呈现正相关性这一特性,可将其作为细胞酪氨酸产量的传感器,为高通量筛选酪氨酸高产菌株提供了思路。  相似文献   

3.
抑制黑色素合成的乳酸菌胞外多糖的筛选和性质研究   总被引:1,自引:0,他引:1  
【目的】筛选可抑制黑色素合成的乳酸菌胞外多糖。【方法】通过观察凝乳拉丝外观筛选产胞外多糖的乳酸菌菌株,测量胞外多糖对B16黑色素瘤细胞黑色素合成和细胞活力的影响。对胞外多糖进行纯化,并通过PMP衍生-HPLC、红外光谱、抑制酪氨酸酶活性、抗氧化能力对其单糖组成和结构、作用机制进行研究。【结果】筛选到一株乳酸菌Lactobacillus rhamnosus HLAB122,发酵产生的胞外多糖在5 g/L浓度下可使B16细胞黑色素产量下降至空白对照的32.7%,且在96 h内对细胞活力无影响。纯化后的多糖由鼠李糖、葡萄糖、半乳糖构成,各单糖摩尔比为1?5.44?5.37。该胞外多糖不抑制酪氨酸酶活力且抗氧化性微弱。【结论】L.rhamnosus HLAB122产生的胞外多糖在个人护理产品中有潜在应用价值。  相似文献   

4.
从重庆不同区域收集土壤,分离到一株高产黑色素的菌株(H4),通过16Sr DNA序列测定,菌株与Streptomyces puniciscabiei序列相似性为99%,该菌产生的胞外黑色素在可见-紫外光区均有吸收,紫外区光吸收强,随着波长的减小,吸收增强,在210 nm左右有1个吸收高峰。采用响应面法对其发酵条件进行了优化,结果表明该菌株在L-酪氨酸的含量为0.73 g/L,p H值为6.05,温度28℃的条件下,菌株黑色素实际产量达2.49 g/L。  相似文献   

5.
酪氨酸酶基因编码的酪氨酸酶是生物体合成黑色素的关键酶。采用比较酪氨酸酶的同源保守结构域氨基酸序列的方法设计引物 ,从苏云金芽胞杆菌 (Bacillusthuringiensis) 4D11中通过PCR扩增得到了包含酪氨酸酶基因的DNA片段。将该片段亚克隆到载体pGEM_7zf上并转入大肠杆菌DH5α ,所得到的转化子在添加了L_酪氨酸的LB培养基中能合成可溶性的黑色素。测定该菌株黑色素的产量和在紫外光照射后的菌体活力 ,结果表明该基因产生的黑色素能在一定程度上保护菌体免受紫外辐射  相似文献   

6.
香灰菌黑色素对真菌生长及多酚氧化酶活性的影响   总被引:2,自引:0,他引:2  
以PDB培养的香灰菌丝体为材料,提取、纯化香灰菌黑色素,并测定香灰菌黑色素对六种真菌生长及对其胞外多酚氧化酶活性的影响.结果表明:香灰菌黑色素对六种真菌的生长没有抑制作用,而对其多酚氧化酶活力有明显的诱导作用.在不同的香灰菌黑色素浓度范围内,六种真菌胞外多酚氧化酶最大酶活的出现时间不同.  相似文献   

7.
野生Bc菌株产黑色素的研究*   总被引:1,自引:0,他引:1  
Bc58是一株野生蜡状芽孢杆菌菌株,经L-酪氨酸诱导后可产生红棕色色素。通过红外光谱及各种化学测定证明该色素与S igm a公司标准黑色素(M elan in)的性质相似。生测结果显示添加Bc58黑色素的B t制剂经紫外照射5 h后的LC50为16.1μg/mL,与未经紫外照射的B t制剂的LC5015.2μg/mL基本相同,而比未添加黑色素的B t制剂经紫外照射后的杀虫毒力高出近1倍。经SDS-PAGE检测表明该黑色素可保护苏云金杆菌晶体蛋白在紫外光下基本不降解,表明Bc58黑色素  相似文献   

8.
表面活性剂对嗜热脂肪芽孢杆菌产高温蛋白酶的影响   总被引:7,自引:0,他引:7  
研究了表面活性剂对嗜热脂肪芽孢杆菌(Bacillusstearothermophilus)WF146产胞外高温蛋白酶的影响。结果表明,表面活性剂Tween80在0.05%~0.1%(体积比)浓度范围内对WF146产酶有一定的促进作用。在培养基中添加0.1%Tween80可使发酵液酶活提高12.7%,Tween20和TritonX100则抑制嗜热脂肪芽孢杆菌WF146产酶。另外,TritonX100抑制嗜热脂肪芽孢杆菌WF146生长,而Tween80和Tween20不抑制其生长。  相似文献   

9.
研究了短小芽孢杆菌(Bacillus pumilus)对盐藻空间诱变株系SZ-05(Dunaliella salina SZ-05)的生物量及β-胡萝卜素积累的影响。结果表明,短小芽孢杆菌显著提高了盐藻SZ-05的生物量和β-胡萝卜素的产量,明显降低了培养体系中的溶解氧和胞外多糖的含量。溶解氧的减少,使得藻细胞的光呼吸作用下降,光合作用速率提高,使藻细胞生物量增加。胞外多糖具有抗氧化作用,胞外多糖的减少可能进一步增加了β-胡萝卜素的合成,从而使β-胡萝卜素在胁迫条件下大幅度增加。  相似文献   

10.
【目的】研究西南不同地区的高山湖泊中可培养细菌的多样性及其产胞外蛋白酶、纤维素酶和胞外多糖的能力。【方法】以西南4个不同地区的高山湖泊:雷波的马湖(LB)、中缅边境的凯邦亚湖(ZM)、沙德的莲花湖(SD)、腾冲的青海湖(TC)的水样为研究对象,利用稀释涂布平板方法对可培养细菌进行分离筛选,然后通过对可培养细菌的生理生化指标和16S r RNA基因序列进行分析,初步确定细菌属别;对分离得到的菌株进行产胞外蛋白酶和纤维素酶活性测定和产胞外多糖能力检测。【结果】从西南地区4个湖泊中共分离筛选得到41株细菌,其中LB 15株、ZM 13株、SD 7株、TC 6株。根据16S r RNA基因序列的系统进化分析,4个地区可培养细菌的组成和丰度存在明显差异,其中LB和ZM的优势菌属是芽孢杆菌属(Bacillus),其次是气单胞菌属(Aeromonas)和假单胞菌属(Pseudomonas),分离的TC菌株全部属于芽孢杆菌属(Bacillus),分离的SD菌株特异性较强。进一步酶活性和胞外多糖检测表明,分离得到的41株细菌中有28株菌的发酵产物具有蛋白酶活性,6株具有纤维素酶活性,17株可产胞外多糖(Exopolysaccharides,EPS)。其中有2株细菌同时产蛋白酶、纤维素酶和胞外多糖,10株细菌同时产蛋白酶和胞外多糖,2株细菌同时产蛋白酶和纤维素酶,1株细菌同时产纤维素酶和胞外多糖。【结论】西南4个高山湖泊中存在丰富的微生物菌种资源,且4个湖泊中筛选的可培养细菌受所处环境的影响大。其中莲花湖由于高海拔和较偏僻等特点,人为干扰小,分离得到的细菌类群与其他湖泊相比明显不同;而马湖、凯邦亚湖和青海湖3个湖泊的海拔相对较低,受人类活动影响较大,分离得到的细菌均较常见。此外高山湖泊中的可培养细菌具有分泌多种胞外活性物质特性,为工业化应用奠定了资源基础,极具更深入的开发和研究价值。  相似文献   

11.
AIMS: To isolate a bacterium that produces high yield of melanin and to examine the effect of this bacterial pigment on the efficacy of a bioinsecticide. METHODS AND RESULTS: A novel melanin-producing bacterium, designated as strain WS, was isolated from the East Lake, Wuhan, China. Taxonomic studies of this strain indicate that it belongs to Aeromonas media. Physicochemical analysis of the pigment produced by strain WS (melanin WS) suggests that it is the authentic 3,4-dihydroxyphenylalanine (DOPA)-melanin. This melanin and that produced by Pseudomonas maltophilia P28 (melanin P28) share many biophysical properties, but the yield of the melanin WS is significantly higher than that of the melanin P28. In addition, the melanin WS appears to be more effective in the protection of a bioinsecticide against ultraviolet (UV) or solar radiation. At the concentration of 10 ppm, the melanin P28 exhibited no photoprotective effect on the bioinsecticide against UV radiation; in contrast, 5 ppm of melanin WS displayed an obvious protective effect. Similarly, the melanin WS displayed more protective effect on the bioinsecticide against solar radiation than the melanin P28 did over a 4-day period, with the effect being more dramatic for the last 2 days. CONCLUSIONS: We have isolated a novel bacterial strain of A. media that produces high levels of melanin. The melanin produced by this strain offers effective photoprotection of a commercial bioinsecticide BTI against UV and solar radiation. SIGNIFICANCE AND IMPACT OF THE STUDY: Our study suggests that the melanin produced by our newly isolated A. media strain has the potential to be used as a general photoprotective agent for bioinsecticides.  相似文献   

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14.
The bacterium Bacillus thuringiensis produces a crystal protein with insecticidal properties; however, crystal proteins can be damaged by ultraviolet (UV) radiation. The aim of this study was to improve the stability of the insecticidal crystal protein (ICP) by constructing a mutant line that expresses high levels of the UV light-protecting pigment, melanin. BMB181, a B. thuringiensis mutant with high melanin production, was obtained after sub-culturing BMB171 for several generations at 42 °C. The melanin yield by BMB181 (without tyrosine supplementation) reached 8.55 mg/ml. The electroporation efficiency of BMB181 reached 106 CFU/μg when a 6.7-kb foreign plasmid was used. Microscopic and SDS-PAGE analyses revealed that ICP (CryIAc10; GenBank: AAA73077.1), which is highly toxic to Lepidoptera, was synthesized efficiently by strain BMB181. The insecticidal properties of a recombinant line derived from strain BMB181, designated BMB32 (cry1Ac10/BMB181), was tested against the cotton bollworm, Helicoverpa armigera. After UV irradiation for 4 h, BMB32 had a half maximal inhibitory concentration value of 1.37 μg/ml, whereas the control line BMB31 (cry1Ac10/BMB171) had a median lethal dose value of 25.85 μg/ml. These results indicate that the B. thuringiensis mutant is a candidate for industrial scale production of light-stable insecticides.  相似文献   

15.
AIMS: A parametric study was conducted to define optimum conditions to achieve high yields in the conversion of tyrosine to eumelanin (EuMel) using recombinant Escherichia coli. METHODS AND RESULTS: Escherichia coli W3110 (pTrcMutmelA) expressing the tyrosinase coding gene from Rhizobium etli and glucose-mineral media were used to transform tyrosine into EuMel. Batch aerobic fermentor cultures were performed to study the effect of temperature, pH and inducer concentration (isopropyl-D-thio-galactopyranoside) on melanin production. Under optimum conditions, 0.1 mmol l(-1) of isopropyl-D-thio-galactopyranoside, temperature of 30 degrees C, and changing pH from 7.0 to 7.5 during the production phase, a 100% conversion of tyrosine into EuMel is obtained. Furthermore, tyrosine feeding allowed us to obtain the highest level (6 g l(-1)) of EuMel produced by recombinant E. coli reported until now. CONCLUSIONS: The most important factors affecting melanin formation and hence influencing the rate and efficiency in the conversion of tyrosine into EuMel in this system, are the temperature and pH. SIGNIFICANCE AND IMPACT OF THE STUDY: Maximum theoretical yield was obtained using a simple culture process and mineral media to convert tyrosine (a medium value compound) into melanin, a high value compound. The process reported here avoids the use of purified tyrosinase, expensive chemical methods or the cumbersome extraction of this polymer from animal or plant tissues.  相似文献   

16.
The effect of various compounds on growth, melanin biosynthesis and cell differentiation was studied in a hyaline (SH25) and a pigmented (SH25B) strain of Microdochium bolleyi. Dark pigment production by the hyaline strain was induced by the presence of DOPA and indole in the medium, both of which are intermediates in the tyrosine pathway of melanin biosynthesis. These substrates also induced conidia and chlamydospore production, structures which were not normally seen in the hyaline strain. The systemic fungicide, tricyclazole, inhibited melanin synthesis in the pigmented strain but not the development of chlamydospore-like cell structures.  相似文献   

17.
为筛选一株产黑色素能力强的菌株并优化其培养条件。通过ITS测序鉴定11株供试菌株,以菌丝生长速度、平板L值等指标筛选出一株产黑能力强的香灰菌,并对其生长所需碳源、氮源、pH等培养条件进行优化。研究表明,11株供试菌株均为香灰菌(Hypoxylon sp.),其中Hp.sp0006菌丝生长速度较快、菌球大且均匀、L值最低,并且发酵液黑色素含量最高。该菌株最优培养条件是,以葡萄糖为碳源、牛肉浸膏为氮源、碳氮比20∶1并添加10 mg维生素B1,黑色素含量可达(1.21±0.17)g/L。香灰菌Hp.sp0006是一株产黑色素较高的菌株,优化后的培养基更有利于黑色素的合成,为香灰菌黑色素的开发利用奠定基础。  相似文献   

18.
Sporothrix schenckii is the etiological agent of sporotrichosis, the main subcutaneous mycosis in Latin America. Melanin is an important virulence factor of S. schenckii, which produces dihydroxynaphthalene melanin (DHN-melanin) in conidia and yeast cells. Additionally, l-dihydroxyphenylalanine (l-DOPA) can be used to enhance melanin production on these structures as well as on hyphae. Some fungi are able to synthesize another type of melanoid pigment, called pyomelanin, as a result of tyrosine catabolism. Since there is no information about tyrosine catabolism in Sporothrix spp., we cultured 73 strains, including representatives of newly described Sporothrix species of medical interest, such as S. brasiliensis, S. schenckii, and S. globosa, in minimal medium with tyrosine. All strains but one were able to produce a melanoid pigment with a negative charge in this culture medium after 9 days of incubation. An S. schenckii DHN-melanin mutant strain also produced pigment in the presence of tyrosine. Further analysis showed that pigment production occurs in both the filamentous and yeast phases, and pigment accumulates in supernatants during stationary-phase growth. Notably, sulcotrione inhibits pigment production. Melanin ghosts of wild-type and DHN mutant strains obtained when the fungus was cultured with tyrosine were similar to melanin ghosts yielded in the absence of the precursor, indicating that this melanin does not polymerize on the fungal cell wall. However, pyomelanin-producing fungal cells were more resistant to nitrogen-derived oxidants and to UV light. In conclusion, at least three species of the Sporothrix complex are able to produce pyomelanin in the presence of tyrosine, and this pigment might be involved in virulence.  相似文献   

19.
Transglutaminases catalyze the cross-linking and amine incorporation of proteins, and are implicated in various biological phenomena. Previously, we found a high molecular mass transglutaminase-inhibitory substance produced by Streptomyces lavendulae Y-200 that appeared to be a melanin substance. Here, we report that synthetic tyrosine melanin inhibited various types of transglutaminases. Tyrosine melanin inhibited tissue-type transglutaminase in a competitive manner with a glutamine substrate, and also inhibited the cross-linking of casein catalyzed by a tissue-type transglutaminase. The melanized hemolymph of the silkworm and melanin solutions prepared from melanin precursors inhibited tissue-type transglutaminase. These results suggested that the melanin substances generally inhibit transglutaminases.  相似文献   

20.
Melanin isolated from the ink sac of cuttle fish (Sepia melanin) is a proposed standard for natural eumelanin. Sepia melanin isolated by a standard protocol was submitted for both elemental analysis and quantitative amino acid analysis. The contribution of the detected amino acids to the elemental composition is subtracted from the total elemental analysis, and the resultant elemental composition reflects the composition of the Sepia melanin backbone chromophore. The assumption is made that, for eumelanins, there is only one nitrogen atom per monomeric unit, and thus, the empirical formula for the average monomeric Sepia melanin backbone chromophore was determined. Three key parameters can be determined for any melanin sample; namely, the molar C/N for the average monomeric unit, the formula weight of the average monomeric unit, and the total percent composition of amino acid residues. Three commonly used melanin preparations, namely, natural Sepia melanin, melanin prepared by the in vitro tyrosinase catalyzed polymerization of tyrosine (tyrosine-enzymatic melanin), and a polymer synthesized by the peroxide oxidative polymerization of tyrosine (tyrosine-chemical melanin), have been subjected to this standard method of characterization. Tyrosine-enzymatic and Sepia melanin are quite similar and tyrosine-chemical melanin is fundamentally different from the other two melanins.  相似文献   

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