兔肉发酵火腿的研制

以兔腿肉为原料,经腌制、发酵,采用L9(3)4正交试验,研制兔肉发酵火腿,测定其pH值、游离氨基酸和三甲胺-氮含量,建立发酵菌株的最佳组合。结果表明当菌株L26、C18、Y163之间的比例为3:3:2,接种量为3×107cfu/g、3×107cfu/g、2×107cfu/g,发酵温度30~35℃时,火腿中的游离氨基酸含量高,品质最好。     

中高温大曲发酵过程微生物消长规律探析

为全面剖析发酵期中高温大曲微生物更迭过程,本文对洋河大曲发酵期各阶段的曲皮和曲心微生物动态消长规律进行了研究。结果表明,曲皮和曲心酵母菌在第1天即达到峰值(10~7cfu/g),受并房和大火前期逐渐升高的曲温影响,曲皮酵母菌从10~5cfu/g下降至10~4cfu/g,大火后期至发酵结束,曲样中鲜少检出酵母菌;曲皮霉菌于主发酵第2天开始大量繁殖,潮火期达到峰值(10~7cfu/g),此时曲心处霉菌仅为10~4cfu/g,这个差距延续至发酵期结束。大火期后,霉菌数量下滑严重,至发酵结束曲皮霉菌仅剩10~4cfu/g;曲皮细菌于发酵第1天即达到顶峰(10~7cfu/g),后续阶段略有下滑,从10~7cfu/g下降至10~6cfu/g,养曲阶段又回升至10~7cfu/g,与曲皮细菌消长规律截然不同,曲心细菌在发酵第2天才有检出,自此至大火前中期,曲心处细菌生长旺盛,达到峰值(10~7cfu/g),大火后期至发酵结束,曲心细菌数略微下降,数量则维持在106cfu/g。     

双歧杆菌发酵胡萝卜汁饮料的研制

采用已耐氧驯化的两歧双歧杆菌与普通乳酸菌共同发酵胡萝卜汁制得发酵饮料。通过正交试验确定的最优发酵条件为:发酵温度39℃,发酵时间9h,胡萝卜汁浓度27％,乳糖添加量1％,双歧杆菌接种量7％,乳酸菌接种量1％。用此条件所得产品中双歧杆菌和乳酸菌活菌数分别达6.5×10~7cfu/mL和1.7×10~8cfu/mL。该产品是色、香、味俱佳的天然营养保健制品。     

一种复合微生物发酵剂对发酵香肠品质的影响

本研究以分离自如皋火腿的木糖葡萄球菌S253及分离自金华火腿的发酵乳杆菌Y_4为发酵剂,研究不同复配比例对发酵香肠组成成分、理化品质及感官品质的影响。结果表明当添加量为10~7cfu/g,Aw值和p H降低速度显著快于对照组(p0.05);硬度和咀嚼性显著高于对照组(p0.05),TBARS值也显著低于其他组(p0.05);木糖葡萄球菌S_(253)和发酵乳杆菌Y4的添加比例为3∶1(v/v)时,产品的a*/b*值显著高于其他组(p0.05),综合感官评分有显著提高(p0.05)。所以不同复配比例的微生物发酵剂对发酵香肠的品质均有显著的影响。综合产品的总体感官评价结果,当木糖葡萄球菌S253和发酵乳杆菌Y4的添加比例为3∶1(v/v),且添加量为10~7cfu/g时,发酵香肠的综合品质最好。     

3-羟基丙醛耐受菌的筛选与3-羟基丙醛抑菌活力的研究

活化的短乳杆菌用含一定甘油浓度(10 g/L)的平板培养基驯化培养后铺上一层含活化E.coli的培养基,30℃双层平板培养,经7个批次筛选,获得了1株在其周围产生明显抑菌透明圈的短乳杆菌菌落,编号为LB7—6;纯化培养筛选出的短乳杆菌,分别在30℃、37℃下进行二次发酵2～8 h,测定发酵液中3-羟基丙醛(3- HPA)的含量。结果显示,较优化发酵条件为37℃微氧静置发酵6h,在此条件下,3-HPA含量为2.18mg/mL,由此计算得3-HPA总得率为0.147g/g,转化率为19.05%;以10~5个/mL E coil为检测菌,采用比浊法实验,结果表明,发酵所得3-HPA最小抑菌浓度(MIC)为2.18×10~(-3)mg/mL;发酵液40℃保存48 h后测定3—HPA的MIC为2.18×10~0mg/mL,说明所产3-HPA抑菌活力有一定程度的稳定性。     

复合菌发酵生产生物饲料工艺条件的研究

探讨复合菌发酵生物饲料发酵条件的优化,采用植物乳杆菌和酵母菌混合发酵的方法,研究菌种比例、装料量、含水量、发酵温度、接种量、腐植酸钠添加量对发酵产物中活菌数的影响。结果表明：生物饲料的最佳发酵工艺条件为: 植物乳杆菌和酵母菌的接种比例为1 ︰ 1,装料量50 g,含水量50%,发酵温度34℃,接种量10%,腐植酸钠添加量2%。在本试验条件下,复合菌生物饲料中含有植物乳杆菌17.59×10⁸ cfu/g,酵母菌8.84×10^8 cfu/g。     

凝结芽孢杆菌工业化发酵培养基初步研究

针对工业生产的需要,采用正交试验设计,研究筛选了饲料添加剂用凝结芽孢杆菌的发酵培养基。优化后的培养基配方为玉米粉15 g/L,豆粕粉10 g/L,(NH_4)_2SO_46 g/L,玉米浆6 g/L,MgSO_4·7H_2O 1 g/L,K_2 HPO_4·3H_2O 2g/L,MnSO_4 0.05 g/L。在pH7.0～7.2和40℃条件下,培养24 h活菌数量达到7.5×10~9 CFU/mL,36 h活菌数量达到9.3×10~9CFU/mL。     

禽肉发酵枣肠的制备及其成熟过程中品质分析

以鹅脯肉、鸡脯肉、鸭脯肉和猪背膘为原料,选取植物乳杆菌(Lactobacillus plantarum)、木糖葡萄球菌(Staphylococcus xylosus)和戊糖片球菌(Pediococcus pentosaceus)作为发酵剂(菌种配比为1∶1∶1),开发发酵枣肠。通过单因素和L_9(3~4)正交实验,确定了在添加鹅脯肉40%、猪背膘10%的基础上,最优工艺参数为:发酵剂添加量10~7cfu/g,发酵时间20 h,发酵温度20℃,鸡脯肉和鸭脯肉添加比例2∶1(添加量50%)。发酵枣肠成熟过程中,水分含量由66.03%降至24.76%,非蛋白氮(Non-protein nitrogen,NPN)含量由0.13%升至0.35%,游离氨基酸含量(Free amino acids,FAA)由842.00 mg/kg升至1878.33 mg/kg。由此开发具有良好品质和风味的禽肉发酵枣肠,同时缩短了产品的成熟时间。     

利用Lactobacillus casei Zhang开发益生菌新鲜干酪

通过添加不同比例Lactobacillus casei Zhang发酵剂制作新鲜干酪,并对其在新鲜干酪中的活力和添加Lactobacillus后新鲜干酪的理化性质进行了研究。结果表明,Lb.casei Zhang在新鲜干酪中具有较高的活力,添加2%、1%、0.5% Lb.casei Zhang发酵剂的干酪中,4℃冷藏开始前,Lb.casei Zhang活菌数分别为2.24×10~8 cfu/g,1.38×10~8 cfu/g,5.55×10~7 cfu/g,4℃冷藏28d后,Lb.casei Zhang存活率分别为99.12%,98.31%,98.61%。在制作过程中和4℃冷藏过程中,与空白组相比,添加Lb.casei Zhang对新鲜干酪的pH值、滴定酸度、蛋白水解活性影响都不显著(P>0.05)。     

毛细管电泳-电化学检测测定荞麦中表儿茶素、芦丁、槲皮素的含量

用毛细管电泳-电化学检测的方法研究了荞麦中黄酮类物质:表儿茶素、芦丁、槲皮素的含量,研究了电极电位、缓冲液的pH值、分离电压及进样时间对电泳的影响,得到优化的测定条件。以直径为300μm的碳圆盘电极为检测电极,工作电极电位为0.95V(vs.SCE),在50mmol/L硼砂(pH8.5)运行缓冲液中,上述3组分在12min内完全分离。表儿茶素、芦丁、槲皮素线性范围分别为5×10-7￣1×10-4g/mL、1×10-6￣1×10-4g/mL、1×10-6￣1×10-4g/mL;检出限分别为1.83×10-7g/mL、2.9×10-8g/mL、1.00×10-7g/mL;3种标样7次平行进样的相对标准偏差(RSD)小于2.5%;回收率表儿茶素100.4%、芦丁98.1%、槲皮素100.1%(n=3)。该法灵敏可靠,结果令人满意。     

多菌发酵对如式香肠营养与风味的影响

以米酒乳杆菌L04、耳式葡萄球菌C131 和德巴利汉逊酵母Y163 为复合发酵剂研制一种新型如式香肠,以不接菌的自然发酵样品为对照,测定其游离氨基酸与脂肪酸、挥发性风味物含量。结果显示：优配方组起始菌数构成比L04:C131:Y163 为1:2:1 的氨基酸总量、必需氨基酸含量分别比对照组提高103%、54%;亚油酸、花生四烯酸分别提高59%、45%;脂肪酸构成饱和脂肪酸(SFA):单不饱和脂肪酸(MUFA):多不饱和脂肪酸(PUFA)之比由1.0:1.1:0.4 优化为1.0:1.2:0.5;醛类、酮类香气物质分别提高62.07%、13.96%,理想效果归于C131 作用以及与其他菌的巧妙搭配。     

发酵剂对兔肉脯游离脂肪酸变化的影响

将兔肉糜中接种植物乳杆菌（Lactobacillus plantarum）、木糖葡萄球菌（Staphylococcus xylosus）、汉逊氏德巴利酵母（Dabaryomyces hansenii）,发酵及烘烤制成兔肉脯,研究3 种不同菌种接种发酵处理对兔肉脯中游离脂肪酸变化的影响,结果表明：总的游离脂肪酸含量均高于非接种菌组,其含量由高到低分别为葡萄球菌处理组＞酵母菌处理组＞植物乳杆菌处理组,说明添加的发酵剂促进了兔肉脯中脂肪的分解,而且它们在兔肉脯中分解产生游离脂肪酸的能力依次递减,各种脂肪酸的含量各有增减,说明不同的发酵剂对其作用不同。进一步研究3 种菌复合发酵处理对兔肉脯中游离脂肪酸影响情况,结果表明其总的游离脂肪酸含量及各种脂肪酸含量均高于单一菌接种组,说明3 种菌的复合发酵分解产生游离脂肪酸的能力较强。     

Glycolytic sequence and respiration of Debaryomyces hansenii as compared to Saccharomyces cerevisiae

The fermentation and respiration activities of Debaryomyces hansenii were compared with those of Saccharomyces cerevisiae grown to stationary phase with high respiratory activity. It was found that: (a) glucose consumption, fermentation and respiration were lower than for S. cerevisiae; (b) fasting produced a much smaller decrease of respiration; (c) glucose consumed and not transformed to ethanol was higher; (d) in S. cerevisiae, full oxygenation prevented ethanol production but this effect was reversed by CCCP, whereas D. hansenii still showed some ethanol production under aerobiosis, which was moderately increased by CCCP. ATP levels were similar in the two yeasts. Levels of glycolytic intermediaries after glucose addition, and enzyme activities, indicated that the main difference and limiting step to explain the lower fermentation of D. hansenii is phosphofructokinase activity. Respiration and fermentation, which are lower in D. hansenii, compete for the re-oxidation of reduced nicotinamide adenine nucleotides; this competition, in turn, seems to play a role in defining the fermentation rates of the two yeasts. The effect of CCCP on glucose consumption and ethanol production also indicates a role of ADP in both the Pasteur and Crabtree effects in S. cerevisiae but not in D. hansenii. D. hansenii shows an alternative oxidase, which in our experiments did not appear to be coupled to the production of ATP.     

DNA typing methods for differentiation of Debaryomyces hansenii strains and other yeasts related to surface ripened cheeses

The discriminative power of ITS-PCR, ITS-PCR RFLP and mitochondrial (mt)-DNA RFLP were evaluated for differentiation of yeasts of importance for surface ripened cheeses. In total 60 isolates were included. Of these, 40 strains of the following species, Debaryomyces hansenii var. hansenii, D. hansenii var. fabryi, Saccharomyces cerevisiae, Candida zeylanoides, Kluyveromyces lactis and Yarrowia lipolytica, were obtained from culture collections and 20 isolates of D. hansenii representing six different phenotypes were collected from seven Danish producers of surface ripened cheeses. ITS-PCR was evaluated for differentiation at species level on the 40 strains obtained from culture collections. Ten strains of each variety of D. hansenii and five strains of each of the above mentioned species were analysed. For each of the investigated species, a specific ITS1-5.8S rDNA-ITS2 region size was observed. Accordingly ITS-PCR was found valuable for differentiation at species level of yeasts of importance for surface ripened cheeses. ITS-PCR RFLP was investigated for the purpose of strain typing of D. hansenii. Ten CBS strains of each variety of D. hansenii were analysed. Only one enzyme (TaqI) out of several investigated (BamHI, DpnI, Fnu4HI, HaeIII, HindIII, HpaII, NlaII, Sau3AI, TaqI) demonstrated genetic diversity within the strains. This enzyme divided the 20 strains in three groups. Sequence analysis of the ITS1-5.8S rDNA-ITS2 region for the type strains of each variety of D. hansenii showed an identity of 99.84%, corresponding to a difference in one basepair. Based on these results, ITS-PCR RFLP was found ineffective for strain typing of D. hansenii. MtDNA RFLP using HaeIII and HpaII was evaluated for strain typing of D. hansenii on the 20 CBS strains of D. hansenii. The CBS strains were divided into 16 groups according to their restriction profiles, which proved the method useful for typing of D. hansenii at subspecies level. The 20 dairy isolates showed a lower genetic variability than the CBS strains as they were divided into eight groups. Cluster analysis of the 20 CBS strains and the 20 dairy isolates based on their mtDNA restriction profiles showed (max. similarity level = 52%) that the dairy isolates only clustered with the CBS strains of D. hansenii var. hansenii. For some of the dairies more than one strain of D. hansenii were found to be involved in the ripening process, indicating that the method could be useful for subspecies typing and investigation of the microbial succession between strains of D. hansenii during the ripening process of surface ripened cheeses.     

微生物发酵剂对发酵风鸭中脂类物质的影响

分别以米酒乳杆菌、木糖葡萄球菌和德汉逊氏酵母为发酵剂生产发酵风鸭,就微生物对发酵风鸭中脂类物质的影响进行研究。研究结果表明,与对照相比,微生物发酵剂使风鸭脂类物质的含量与组成发生了显著的变化,游离脂肪酸的含量也显著增加。米酒乳杆菌与德汉逊氏酵母分解脂类物质生成游离脂肪酸的能力较强,而木糖葡萄球菌有较强的分解游离脂肪酸的能力。微生物发酵剂对发酵风鸭中脂类物质的影响是显著的。     

Effects of salts on Debaryomyces hansenii and Saccharomyces cerevisiae under stress conditions

The effect of Na+ and K+ on growth and thermal death of Debaryomyces hansenii and Saccharomyces cerevisiae were compared under stress conditions as those commonly found in food environments. At the supraoptimal temperature of 34 degrees C both cations at concentrations of 0.5 M stimulated growth of D. hansenii, while K+ had no effect and Na+ inhibited growth of S. cerevisiae. At 8 degrees C, close to the minimum temperature for growth in both species, both cations inhibited both yeasts, this effect being more pronounced with Na+ in S. cerevisiae. At extreme pH values (7.8 and 3.5) both cations at concentrations of 0.25 M stimulated D. hansenii while Na+ inhibited S. cerevisiae. K+ inhibited this yeast at pH 3.5. Thermal inactivation rates, measured at 38 degrees C in D. hansenii and at 48 degrees C in S. cerevisiae, decreased in the presence of both cations. This protective effect could be observed in a wider range of concentrations in D. hansenii. These results call the attention to the fact that not all yeasts have the same behaviour on what concerns synergy or antagonism of salt together with other stress factors and should be taken into consideration in the establishment of food preservation procedures.     

Expression of the GPD1 and GPP2 orthologues and glycerol retention during growth of Debaryomyces hansenii at high NaCl concentrations

The highly NaCl-tolerant yeast Debaryomyces hansenii produces and obtains high levels of intracellular glycerol as a compatible solute when grown at high NaCl concentrations. The effect of high NaCl concentrations (4%, 8% and 12% w/v) on the glycerol production and the levels of intra- and extracellular glycerol was determined for two D. hansenii strains with different NaCl tolerance and compared to one strain of the moderately NaCl-tolerant yeast Saccharomyces cerevisiae. Initially, high NaCl tolerance seems to be determined by enhanced glycerol production, due to an increased expression of DhGPD1 and DhGPP2 (AL436338) in D. hansenii and GPD1 and GPP2 in S. cerevisiae; however, the ability to obtain high levels of intracellular glycerol seems to be more important. The two D. hansenii strains had higher levels of intracellular glycerol than the S. cerevisiae strain and were able to obtain high levels of intracellular glycerol, even at very high NaCl concentrations, indicating the presence of, for example, a type of closing channel, as previously described for other yeast species.     

The co-action of osmotic and high temperature stresses results in a growth improvement of Debaryomyces hansenii cells

Debaryomyces hansenii is a salt tolerant yeast species, often isolated from sea water or found among other spoilage yeasts in several types of food. In this work, we examined the influence of temperature and increased osmotic pressure (two parameters also important in food industry) on D. hansenii growth. Several other authors showed that its growth at the normal yeast cultivation temperature (28 to 30 degrees C) is stimulated by the presence of sodium, in contrast to the growth of Saccharomyces cerevisiae, which is inhibited by the presence of sodium under the same experimental conditions. Here we show that the previously reported growth stimulation by sodium is temperature dependent in D. hansenii and can be observed under conditions that already amount to high temperature stress for D. hansenii. At a lower temperature (more convenient for D. hansenii cultivation), we found no significant improvement or even an inhibition of cell growth in the presence of Na(+). The growth of D. hansenii at high temperatures is also improved by the presence of potassium or sorbitol. Moreover, the temperature dependence of stimulatory effects of increased osmotic pressure in media does not seem to be unique for D. hansenii; similar relationships between the growth, cultivation temperature and presence of osmolytes we also observed for S. cerevisiae and Schizosaccharomyces pombe.     

A beta-glucuronidase-based agar medium for the differential detection of the yeast Debaryomyces hansenii from foods

A selective and differential solid medium, Debaryomyces differential medium (DDM), was used for the isolation of Debaryomyces hansenii. This medium is formulated to allow detection of the beta-glucuronidase enzyme using the chromogenic substrate magenta-glucuro.CHA (5Br-6Cl-3indolyl-beta-D-glucuronide, cyclohexylammonium salt). Of the more than 120 microorganisms tested, including yeasts, bacteria, and a filamentous fungus, only D. hansenii produced violet colonies, thus permitting its easy discrimination from other organisms. When quality assessment tests were performed, optimal productivity and selectivity were obtained for D. hansenii. The medium was also satisfactory when used to test naturally contaminated food products.     

Behavior of Brevibacterium linens and Debaryomyces hansenii as ripening flora in controlled production of smear soft cheese from reconstituted milk: growth and substrate consumption dairy foods

Experimental cheeses inoculated with Debaryomyces hansenii and Brevibacterium linens were ripened for 76 d under aseptic conditions. Triplicate cheese-making trials were similar as a result of efficient control of the atmosphere. In all trials, D. hansenii grew rapidly during the first 2 d and then slowed, but growth remained exponential until d 10 (generation time around 70 h). Total cell counts were higher than the number of viable cells, and after 10 d they remained around 3 x 10(9) yeast/g of DM. This difference resulted from the nonviability of a fraction of D. hansenii. After d 15, the pH of the rind was close to 7, and B. linens grew exponentially until d 25 (generation time around 70 h). The growth rate subsequently decreased but remained exponential (generation time around 21 d). Cell counts of D. hansenii and B. linens were correlated with the environmental technical conditions. Total D. hansenii counts were also correlated with total B. linens counts. Viable B. linens counts were related to rind lactate, and total counts depended on rind pH, internal lactate, and D. hansenii viable counts. The internal pH of the cheese depended on lactate concentrations, whereas surface pH was related to internal lactose, temperature, and relative humidity. These results suggest a determining role of the diffusion of the carbon sources in the ripening of smear soft cheese.