Statistical modeling and optimization for exopolysaccharide production by Lactobacillus confusus in submerged fermentation under high salinity stress

In the last two decades, many studies have been reported that a high concentration of NaCl suppresses exopolysaccharide (EPS) production in lactic acid bacteria. In the present study, however, the enhancement of EPS production by Lactobacillus confusus under high salinity stress in submerged fermentation was demonstrated using response surface methodology via a full factorial design. Under the optimized conditions of 3.33% NaCl, 20 g/L sucrose, and 35 h of incubation, the EPS yield was 10.87 g/ L with 178% higher than the maximum yield (6.12 g/L of EPS) produced from the modified MRS medium without NaCl. Biomass production was independent of EPS production. A high yield of biomass was obtained in the culture with 0.23% NaCl. This results indicate that high salinity stress by NaCl can enhance EPS production in submerged fermentation in uncontrolled pH cultivations by inducing the production of cell-associated dextransucrase.     

盐度和加曲量对罗非鱼片加工废弃物速酿鱼露的影响

对罗非鱼片加工废弃物速酿鱼露的发酵工艺进行了初步研究，从氨基酸态氮、总酸、TVBN、色度、感官5个指标分析了速酿鱼露在不同盐度（4，7，10，13g／dL）和不同加曲量（质量分数10％、15％、2S％、35%）下的发酵状况，综合各指标的影响确定适宜的速酿发酵条件为：7g／dL盐度和10％的加曲量，在此条件下的鱼露发酵液氨基酸态氮、总酸含量较高、腐败程度低，风味感官评价较好。     

烟草NtSnRK2.1基因的克隆及其在非生物胁迫

植物SnRK2基因家族(蔗糖非发酵相关蛋白激酶家族2, Sucrose non-fermenting-1-related protein kinase 2)在糖代谢和抗逆信息传递途径中发挥重要作用。本研究从普通烟草腺毛cDNA文库中筛查烟草SnRK2基因的EST序列,以此序列为信息探针,通过电子克隆和RT-PCR的方法从烟草中克隆到一个包含1017 bp开放阅读框、编码338个氨基酸的cDNA序列,命名为NtSnRK2.1。生物信息学分析表明,NtSnRK2.1蛋白同时具有磷酸化丝氨酸/苏氨酸和酪氨酸的活性。氨基酸序列比对发现,NtSnRK2.1与拟南芥、水稻和小麦等植物中受逆境胁迫诱导表达的直系同源基因高度同源。组织表达分析结果显示,烟草NtSnRK2.1基因的组织表达差异较大,在根部的表达量最高,其次是叶片,茎部的表达量最低。胁迫处理下的基因表达分析结果表明,NtSnRK2.1受高盐、高渗、低温胁迫和ABA处理诱导表达,对各胁迫条件的应答模式不同,其对各胁迫条件的敏感度为:高渗＞高盐＞低温＞ABA。      

Salt effect on phenolics and antioxidant activities of Tunisian and Canadian sweet marjoram (Origanum majorana L.) shoots

BACKGROUND: Two varieties of Origanum majorana (Canadian and Tunisian) were evaluated for their phenolic, flavonoid and tannin contents, individual phenolic compounds and antioxidant activities under NaCl constraint. RESULTS: The results showed a significant variability in phenolic composition and antioxidant behavior between the two varieties under salt stress. The phenolic composition of methanolic extracts was determined by reversed‐phase high‐performance liquid chromatography. Amentoflavone was the predominant flavonoid compound; in addition, trans‐2‐hydrocinnamic acid became the major phenolic acid with salt treatment of the Tunisian variety. In the control, Canadian variety extract was characterized by high levels of gallic acid and amentoflavone. However, under 75 mmol L^?1 NaCl, gallic acid content doubled, whereas amentoflavone content was maintained in the Canadian variety. Stimulation of phenolic acid biosynthesis was observed in these two varieties under salt treatment despite the fact that shoots of the Tunisian variety showed higher antioxidant activities compared to those from the Canadian variety. Tunisian O. majorana might have developed tolerance to salinity and avoided tissue damage by activating enzymes involved in the galactosylation of quercetin into quercetin‐3‐galactoside and quercetin‐3‐rhamnoside. CONCLUSION: Our results confirmed the tolerance of Tunisian O. majorana plants. Copyright © 2012 Society of Chemical Industry     

Salt shock enhances the expression of ZrATP2, the gene for the mitochondrial ATPase beta subunit of Zygosaccharomyces rouxii

In the course of a study of cell wall proteins from the salt-tolerant yeast Zygosaccharomyces rouxii, a protein that increased its expression as the NaCl concentration of the culture medium increased was identified. Several degenerate primers were constructed based on partial amino acid sequences of this protein and were used in PCR amplification of a gene termed ZrATP2. The amino acid sequence deduced from nucleotide sequence of the gene revealed that ZrATP2 encodes the beta subunit of mitochondrial F1 ATPase. Northern blot analysis demonstrated that NaCl shock induced an elevation in ZrATP2 expression, which corresponded with the resumption of Z. rouxii cell growth after salt shock.     

发酵盐浓度对Chlorella vulgaris生物合成蛋白与油脂的影响

实验室条件下,考察了发酵过程中不同盐度对小球藻细胞生长、蛋白及油脂含量的影响,结果表明:培养时间为144h,小球藻在无NaCl培养基中生长最佳,其细胞干重、光合色素含量、比生长速率和最大生产强度分别为0.750g·L-1、39.50mg·g-1、0.225d-1和0.125g·L-1·d-1,且氮磷利用率高分别为96.4%及93.5%,其消耗效率分别为9.10、0.71mg·L-1·d-1,藻蛋白含量也最高为55.0%（W）,其生产效率为68.40mg·L-1·d-1;而藻细胞培养在2.5%NaCl浓度下,在96h脯氨酸含量达最高为2.25%（W）,最高与最低值之比超过20.0;小球藻培养在含2.5%NaCl培养基中,在144h油脂含量最高达15.5%（W）,其生产效率为16.10mg·L-1·d-1。NaCl浓度不仅影响小球藻细胞对氮磷等营养的吸收,而且对生物量、蛋白质、脯氨酸及油脂等细胞组分的生物合成具有明显影响。藉此,提出了通过改变盐浓度,达到调控小球藻胞内生理代谢组分的可行性。      

嗜盐性蛋白酶产生菌Virgibacillus sp. P-4的筛选鉴定及其特性分析

利用酪蛋白琼脂平板从传统虾酱中分离嗜盐性蛋白酶产生菌,经形态学、生理生化和分子生物学鉴定,确定该菌隶属于枝芽孢杆菌属Virgibacillus sp.P-4,并用此菌为后续研究菌株。结果表明,该菌最适生长温度为30~37℃,最适生长NaCl质量分数为5%~15%;所产胞外蛋白酶在20~50℃条件下酶活力较高且保持稳定,最适反应温度为40℃,当NaCl质量分数为15%时酶活力达到最高,并且所产胞外蛋白酶种类可能不只一种,但不包含巯基蛋白酶;通过分析酶解液中游离氨基酸释放速率,预测其酶切位点为Phe-、Tyr-、Lys-、His-、Pro-及Leu-。     

Characterization of the low-salinity stress in Vibrio vulnificus

Vibrio vulnificus is a marine pathogenic bacterium commonly found in seawater or seafood. This organism encounters low-salinity stress in its natural environment and during food processing. This study was designed to investigate the response of V. vulnificus YJ03 to lethal low salinity (0.04% NaCl) and its adaptation to sublethal salinity (0.12% NaCl with 20 amino acids added). A short period in the nonculturable state was induced by lethal low-salinity stress followed by cell death after 30 min of stress. Addition of 1 mM glycine betaine or 0.5 mM sucrose reduced the damage. Low-salinity adaptation was achieved in the exponential-phase cells but not in the stationary-phase cells. Significant protection against lethal low-salinity stress was attained when the cells were adapted for as little as 1.5 min. The adapted cells were significantly protected against lethal low salinity and 2.4% sodium sorbate but sensitized to the challenge of heat (52 degrees C) and acid (pH 3.2). Nonlethal low-salinity treatment of seafood should be avoided to prevent stress adaptation of V. vulnificus.     

大豆发芽富集γ-氨基丁酸的培养液组分优化及盐胁迫下的富集机理

以东北大豆为原料,研究培养液组分对大豆发芽富集γ-氨基丁酸(γ-aminobutyric acid,GABA)的影响,利用响应面法优化了大豆发芽富集GABA的培养液组分,在此基础上对低盐胁迫下大豆发芽富集GABA的机理进行研究。结果表明:优化后有效的培养液组分为谷氨酸钠1.0 mg/mL、磷酸吡哆醛2.0 mmol/L、CaCl_2 2.0 mmol/L、NaCl 100 mmol/L,在此条件下,富集得到的发芽大豆中GABA含量较高,为(269.93±4.73)mg/100 g,比大豆发芽前提高了约10倍;盐胁迫下,发芽大豆谷氨酸脱羧酶(glutamate decarboxylase,GAD)活性和GABA含量随Na Cl浓度加大和胁迫时间延长而提高,同时大豆发芽期间GABA含量与其他指标之间相关性分析表明,盐胁迫下发芽大豆GABA含量与芽长、GAD活性、游离氨基酸和可溶性蛋白含量之间呈显著正相关,在低盐胁迫下,大豆发芽受到抑制,但促进了GAD活性的升高,游离氨基酸和可溶性蛋白质含量增加,富集产生了较多的GABA。     

云南鲷鱼骨胶原蛋白的制备及其理化性质

以云南鲷鱼骨为原料,采用低温酸法提取制备鱼骨中的酸溶性胶原蛋白（acid-soluble collagen,ASC）, 并对ASC的氨基酸组成、亚基组成、红外吸收、紫外吸收、热稳定性、X射线衍射、微观结构、多肽片段以及溶解 性进行了全面的分析。氨基酸组成表明ASC主要含甘氨酸、脯氨酸和丙氨酸,而酪氨酸、蛋氨酸和半胱氨酸含量 较低;十二烷基硫酸钠-聚丙烯酰胺凝胶电泳结果显示ASC为Ⅰ型胶原蛋白;ASC在230 nm波长处有最大紫外吸收 峰;傅里叶变换红外光谱和X射线衍射图谱表明ASC分子排列紧凑,保持了其原有的三螺旋结构;差示扫描量热分 析结果显示ASC的变性温度分别为86.5 ℃和226.2 ℃,有较好的热稳定性;扫描电子显微镜显示ASC分子分布均匀、 表面光滑呈三维立体结构;多肽片段分析结果显示鱼骨胶原的氨基酸构成主要为Gly-X-Y,符合胶原蛋白的一级结构 的特点;在pH值小于4的条件下,ASC溶解度较高,当NaCl质量分数大于4%时,ASC溶解度剧烈下降。     

盐胁迫对不同时期盐地碱蓬组分含量的影响

采用自吸水耐盐鉴定法,研究了不同NaCl浓度（0.10%、0.30%、0.60%、0.90%、1.20%）对盐地碱蓬不同生长时期组分含量的影响。结果表明,不同生长时期的盐地碱蓬组分含量存在明显差异,其中苗期盐分对亚硝酸盐影响最大,最高亚硝酸盐含量为7.09 mg/kg;花期盐分对氨基酸含量影响最大,最高含量达134.26 μmol/g;结果期盐分对蛋白质影响最大,最高含量为19.36 mg/g。苗期盐地碱蓬水分含量最高,随盐胁迫程度增加而增加,在1.20% NaCl处理下达最大值;抗坏血酸在不同时期随梯度盐分增加呈现总体降低趋势,低浓度（小于0.60%）NaCl处理下,苗期盐地碱蓬抗坏血酸含量最高。以总黄酮含量为优化目标,采用高效液相色谱检测方法（HPLC）对盐地碱蓬总黄酮提取技术进行了优化,得到最优提取条件:超声功率500 W,超声温度70 ℃,超声时间25 min,料液比1:60 g/mL;在0.90% NaCl处理下,花期盐地碱蓬总黄酮含量最高,达3.68%。综上,本研究明确了适宜发展优质安全盐地碱蓬的土壤盐分范围为0.60%,为指导盐地碱蓬生产和合理开发提供技术支撑和理论依据。     

Effects of hypoxia/hypercapnia on the metablism of Callosobruchus chinensis (L.) larvae

Insects that are pests of stored products are highly tolerant to low O₂ levels (hypoxia) and high carbon dioxide levels (hypercapnia) for long periods, but the underlying mechanism remains unclear. In the present study, we investigated the metabolic profiles of Callosobruchus chinensis (L.) larvae held under 2% O₂+18% CO₂ (both v/v) for 24, 48, and 96 h using gas chromatography–time-of-flight mass spectrometry (GC-TOF-MS), their corresponding control groups under normoxia were also investigated simultaneously. We found that the relative quantity of 91, 24, and 48 metabolites were altered at these three time periods, respectively. The metabolites were principally carbohydrates, organic acids, amino acids, and free fatty acids, 30 of these metabolites with high accuracy of qualitative identification and biological functions were chosen for further analyses. These were intermediates of the citric acid cycle, glycolysis, fatty acid synthesis, and amino acid metabolism. Based on our results, levels of organic acids increased, while levels of amino acids decreased, suggesting that metabolism was shifting from aerobic to anaerobic. Lipid metabolism may be relatively less important; the number of fatty acids exhibiting significant changes in level was quite small. The metabolic profiles changed dynamically over time, with the largest changes evident for 24 h of hypoxia/hypercapnia, followed by 96 h, and only a little change has been shown on 48 has revealed by heatmap and pathway topological analysis. Therefore, we present direct evidence that the metabolism of insect pests of stored products is influenced by complex environmental stressors.     

不同电渗析条件下酱油主要风味组分迁移规律研究

采用电渗析法对酱油进行脱盐处理时，可回收大量具有特殊风味的盐液。该实验研究了不同电渗析条件下脱盐对酱油各组分迁移情况的影响，并分析了盐液的风味物质构成。结果表明，电渗析电压的变化对酱油中氯化钠（NaCl）迁移效率的影响远大于氨基酸态氮及总酸。在电渗析电压13 V、处理时间30 min条件下，酱油的氨基酸态氮（AAN）及总酸（TA）的损失率较低，且NaCl脱除率达41.97%，盐液中NaCl、AAN、TA含量分别为6.60 g/100 mL、0.135 g/100 mL、0.208 g/100 mL。盐液中含有以丙氨酸、亮氨酸及赖氨酸为主的15种游离氨基酸（FAA），总游离氨基酸含量为0.31%。其中鲜味氨基酸的迁移率较低，甜味及苦味氨基酸的迁移率较高。盐液中初步检测到17种风味物质，其中包含苯乙醇、3-甲硫基丙醇、3-甲基-1-丁醇、糠醇、4-乙基愈创木酚等多种酱油特征性风味物质。     

Susceptibility of the heat-, acid-, and bile-adapted Vibrio vulnificus to lethal low-salinity stress

As a marine pathogenic bacterium that inhabits seawater or seafood, Vibrio vulnificus encounters low salinity and other stresses in the natural environment and during food processing. This investigation explores the cross-protective response of sublethal heat-, acid-, or bile-adapted V. vulnificus YJ03 against lethal low-salinity stress. Experimental results reveal that the acid (pH 4.4)- and heat (41 degrees C)-adapted V. vulnificus were not cross-protected against the lethal low-salinity challenge (0.04% NaCl). The bile (0.05%)-adapted exponential- and stationary-phase cells were cross-protected against low salinity, whereas low-salinity (0.12% NaCl)-adapted stationary cells were sensitized against 12% bile stress. Results of this study provide further insight into the interaction between low salinity and other common stresses in V. vulnificus.     

Candida etchellsii产2(5)-乙基-4-羟基-5(2)-甲基-3(2H)-呋喃酮的影响因素

在埃切假丝酵母(Candida etchellsii)发酵过程中,通过盐度调控和氨基酸添加,强化目标产物HEMF(2(5)-乙基-4-羟基-5(2)-甲基-3(2H)-呋喃酮)的合成效率。分阶段调控发酵体系的盐度(初始阶段控制CaCl浓度为200 g/L,发酵40 h后提升至220 g/L),结合氨基酸添加(向发酵体系中添加丙氨酸、精氨酸和甘氨酸各1.67 g/L)。摇瓶结果表明:酵母C.etchellsii合成HEMF,其产量为110.74 mg/L。7 L发酵罐上罐验证,HEMF产量达到121.51 mg/L,相比空白(200 g/L CaCl浓度下且没有添加氨基酸)提高了21.2倍。分阶段盐度调控结合氨基酸添加策略显著强化了C.etchellsii对HEMF的合成。     

Characterization of low salinity stress in Vibrio parahaemolyticus

Vibrio parahaemolyticus is a marine foodborne pathogenic bacterium commonly found in seawater or seafood. This bacterium often encounters low salinity stress when the contaminated seafood is washed with fresh water during food processing. This study was conducted to investigate the response of exponential- and stationary-phase cells of V. parahaemolyticus ST550 to lethal or sublethal low salinity. Tolerance to lethal low salinity (0.25% NaCl) was enhanced in V. parahaemolyticus cells in the exponential phase by previous adaptation in sublethal low salinity (0.6% NaCl). Low salinity-adapted cells in the exponential phase were also cross-protected against the challenge of lethal low pH, indifferent to heat, and sensitized to bile, acetic acid, and lactic acid stress. The adapted cells in the stationary phase were significantly protected against heat treatment at 44°C for 10 and 15 min, sensitized to bile and acetic acid treatment, and indifferent to low pH and lactic acid.