Simultaneous saccharification and fermentation of furfural residues by mixed cultures of lactic acid bacteria and yeast to produce lactic acid and ethanol

In a cellulosic ethanol production system, yeasts cannot be reused, and it is difficult to avoid the formation of lactic acid. A novel mixed fermentation system based on water-rinsed furfural residue was designed to produce lactic acid and ethanol simultaneously by yeast and lactic acid bacteria. The fermentation broth can be used for the production of lactic acid, ethanol, or ethyl lactate, which is a very suitable alternative green solvent. Simultaneous saccharification and fermentation by mixed cultures of lactic acid bacteria and yeast (MSSF) in different conditions were carried out. Acid/alcohol molar ratios (molar ratio of lactic acid to ethanol) were investigated to determine the effects of temperature, substrate concentration, and mass rate of yeast to lactic acid bacterial cells on MSSF. Cellulose conversion rate was also calculated to evaluate the effectiveness of MSSF. The cellulose conversion rate of MSSF was higher than those of ethanol simultaneous saccharification and fermentation and lactic acid simultaneous saccharification and fermentation. The cellulose conversion rate increased with increasing substrate concentration, while the acid/alcohol molar ratio decreased with increasing substrate concentration. These results indicate that yeast cells could provide nutrients for lactic acid bacteria. MSSF at 38 °C is also apt to obtain a low acid/alcohol molar ratio. A 1:1 lactic acid/alcohol molar ratio will be obtained at a fluctuating temperature (38 °C between 0 and 54 h, 42 °C after 54 h) and a substrate concentration of 9%, while keeping a high cellulose conversion rate and final lactic acid concentration. A probable bottleneck for MSSF and its potential solution are also proposed in this paper.     

奶啤生产中乳酸菌对酵母菌发酵作用的研究

利用乳酸菌和酵母菌的共生作用,对牛乳进行了混合菌种的发酵研究。分析了共生作用对产品pH值、滴定酸度、乙醇含量和α-氨基酸态氮的影响。研究结果表明,乳酸菌的加入不仅可以提高酵母菌的生长速度,而且可以提高产品的风味,奶啤的酒精含量相对啤酒较低,是酒精度较低的健康型饮料。     

发酵条件对酸樱桃发酵汁挥发性风味物质的影响探究

目的 考察发酵条件对酸樱桃发酵汁中风味物质的影响。方法 利用3株乳酸菌发酵酸樱桃汁, 以pH为指标明确最佳发酵菌种和稀释比例。随后利用气相色谱-离子迁移谱法(gas chromatography-ion mobility spectrometry, GC-IMS)探究糖添加量、pH、氮源、稀释比例对发酵挥发性风味物质的影响。结果 最佳发酵菌为鼠李糖乳杆菌(Lactobacillus GG, LGG), 最佳稀释比例为1:3, 氮源对挥发性风味物质几乎没有影响; 而不同添加量的碳源(葡萄糖)导致发酵液乙偶姻、双乙酰等含量差异; 随着pH增加, 乙偶姻、双乙酰、2-戊酮等含量显著升高; 稀释比例通过影响发酵原汁的浓度导致发酵后风味物质乙偶姻含量显著增加、2-戊酮含量显著降低。进一步差异比对3株乳酸菌酸樱桃发酵汁的风味, 发现LGG代谢脂肪酸可产生较多的2-庚酮及2-戊酮。结论 菌种、糖添加量、稀释比例、起始pH均可导致发酵液中乙偶姻、2-庚酮及2-戊酮等挥发性风味物质产生显著影响, 本研究为酸樱桃的加工利用提供理论指导。     

Microbiological changes and their impact on quality characteristics of red hot chilli pepper mash during natural fermentation

Production of hot sauce may require fermentation of red hot pepper mash in barrels from 2 weeks up to 3 years. The purpose of this study was to evaluate the microbiological changes and their impact on quality characteristics of red hot pepper mash during natural fermentation over a period of 18 months. Aerobic plate count (APC), lactic acid bacteria (LAB) and yeast counts, as well as changes in pH, acid content, colour and aroma of red hot pepper mash, were analysed. LAB and yeast presented a symbiotic association throughout the fermentation process. Significant pH reduction was observed, with an inverse correlation with acid content. There were minor changes in the colour of the mash. Aroma was analysed based on six volatile compounds which had a significant increase during the first 60 days of fermentation, followed by a significant reduction after 300 days.     

响应面法优化马奶酒发酵条件的研究

以新鲜马奶为原料,通过考察接菌量(乳酸菌/酵母菌)、发酵温度、发酵时间、糖添加量单因素试验对马奶酒发酵的影响,并借助响应面设计,研究接菌量(乳酸菌/酵母菌)、发酵温度、发酵时间对马奶酒品质的影响。结果表明,最佳发酵条件分别为接菌量(乳酸菌/酵母菌)3%∶5%、发酵温度32℃、发酵时间80h、糖添加量6%。马奶酒酒香清雅,微酸爽口。     

Diversity of bacteria and yeast in the naturally fermented cotton seed and rice beverage produced by Brazilian Amerindians

Microorganisms associated with the fermentation of cotton seed and rice were studied using a combination of culture-dependent and -independent methods. Samples of the cotton seed and rice beverage were collected every 8 h during the fermentation process for analysis of the microbiota present over 48 h. The lactic acid bacteria (LAB) population reached values of approximately 8.0 log cfu/mL. A total of 162 bacteria and 81 yeast isolates were identified using polyphasic methods. LAB (Lactobacillus plantarum, Lactobacillus vermiforme, Lactobacillus paracasei) were the most frequently isolated bacteria. Bacillus subtilis was present from 16 h until the end of the fermentation process. A decrease in pH value from 6.92 (0 h) to 4.76 (48 h) was observed, and the concentration of lactic acid reached 24 g/L at the end of the fermentation process. DGGE (denaturing gradient gel electrophoresis) was performed to determine the dynamics of the communities of bacteria and yeast, and the analysis revealed a predominance of LAB throughout the fermentation process. No changes were observed in the yeast community. The yeast species detected were Candida parapsilosis, Candida orthopsilosis, Clavispora lusitaniae and Rhodotorula mucilaginosa. Our studies indicate that the DGGE technique combined with a culture-dependent method is required to discern the dynamics in the fermentation of cotton seed and rice.     

苗族白酸汤中优势乳酸菌及酵母菌的筛选与应用

目的:为了得到适合白酸汤发酵的发酵剂,实现传统白酸汤的规范性生产。方法:采用溶钙圈法结合总酸产量和耐酸耐胆盐实验筛选乳酸菌优良菌株;采用糖苷酶活性实验筛选酵母菌优良菌株;通过生理生化实验、16S rDNA序列和ITS序列进行菌株鉴定。将两株优良发酵菌株复配应用于白酸汤中,对其pH、总酸、总糖、还原糖和感官品质进行分析。结果:从128株乳酸菌中筛选出1株产酸能力较强的菌株G1-3,该菌株在pH3.0与0.3%牛胆盐环境中均生长良好,从20株酵母菌中筛选出1株糖苷酶活性较强的菌株G2-2。经鉴定菌株G1-3为副干酪乳酸杆菌(Lactobacillus paracasei),G2-2为印迪卡有孢圆酵母(Torulaspora indica)。两株菌复配发酵后的籼米白酸汤的总酸含量较自然发酵提高了5倍,总糖与还原糖含量分别降低了61.53%与81.02%;糯米白酸汤中的总酸含量较自然发酵提高了18倍,总糖与还原糖含量分别降低了68.86%与86.99%。两株菌复配发酵后的白酸汤的感官品质也优于自然发酵。结论:G1-3与G2-2的复合发酵剂发酵得到的白酸汤具有较好的品质,有望应用于白酸汤的发酵生...     

酸笋中高产乳酸乳酸菌的筛选、鉴定及发酵条件优化

为获得酸笋中高产乳酸乳酸菌,以柳州传统发酵酸笋为原料,利用MRS培养基对乳酸菌进行分离纯化,然后通过溶钙圈法和高效液相色谱（HPLC）法对高产乳酸乳酸菌进行筛选,并采用形态观察及分子生物学技术对其进行菌种鉴定,最后优化该菌种产乳酸的发酵条件。结果表明,分离并筛选得到一株高产乳酸的乳酸菌菌株LB-1-23,并鉴定其为植物乳杆菌（Lactobacillus plantarum）,该菌株发酵产乳酸的最适条件为接种量2.6%,pH值5.5,发酵温度32 ℃,发酵时间30 h,葡萄糖为碳源,细菌学蛋白胨为氮源。在此优化条件下,乳酸产量为12.74 g/L。该研究为酸笋来源乳酸菌发酵产生乳酸的应用奠定了理论基础。     

West African sorghum beer fermented with Lactobacillus delbrueckii and Saccharomyces cerevisiae: shelf‐life and consumer acceptance

The fermentation profiles, shelf‐life and consumer acceptance of traditional West African sour sorghum beer (pito) fermented with pure commercial starter cultures of lactic acid bacteria (L. delbrueckii) and yeast (Saccharomyces cerevisiae) were evaluated. The beers from this ‘pure culture’ approach were compared with the spontaneous fermentation of pito wort. Lactic acid formation, pH change and extract utilisation were monitored during fermentation. Lactic acid content was used as a measure of sourness to establish the spoilage level of over‐sourness. Further, regression models relating sourness to the time the drink was kept were used to predict the shelf‐life. Consumer acceptance of the product was evaluated using a novel nine‐point hedonic scale. The pure culture and traditionally fermented beers followed similar lactic acid and fermentation profiles but strain‐specific differences were observed. Similar levels of pH, lactic acid level and extract utilisation were achieved. An improvement in shelf‐life of two days was found over traditionally fermented pito. There was no statistical difference between the two pito products for overall liking and taste. However, there was a preference for the aroma of the pure culture pito. It is suggested that the use of pure cultures will facilitate the scale‐up of pito production. © 2019 The Institute of Brewing & Distilling     

乳酸菌与酵母菌对牦牛酸乳品质形成的影响

为明确乳酸菌和酵母菌在牦牛酸乳品质形成中发挥的作用,以牦牛乳粉为原料、牦牛酸乳为发酵剂发酵酸乳,在发酵过程中分别抑制乳酸菌和酵母菌的活性,测定并比较正常发酵、抑制乳酸菌发酵、抑制酵母菌发酵酸乳的风味、口感、质构等相关指标。结果表明,乳酸菌和酵母菌对牦牛酸乳品质形成均起到关键作用,乳酸菌对酸乳酸度、质构、氨基酸态氮、有机酸、V_(B1)及酮类和酸类风味物质的贡献较大,酵母菌对乙醇、V_(B2)及醇类和酯类风味物质的贡献较大。     

自然与乳酸菌强化发酵过程中红酸汤品质变化

利用红酸汤为原料,以总酸、亚硝酸盐、pH、还原糖、可溶性固形物、水分、抗坏血酸为检测指标,分析不同发酵时间自然与乳酸菌强化发酵的红酸汤品质变化规律。结果表明,在整个发酵过程中,自然与乳酸菌强化发酵的红酸汤中还原糖、抗坏血酸、可溶性固形物均呈逐渐下降的趋势;总酸含量和pH均在第6天达到峰值,分别为10.11 g/kg和3.04与10.26 g/kg和2.84;水分含量在第6天达到最低,分别为84.83%与84.18%;亚硝酸盐含量最高分别为0.052 2 mg/kg与0.030 4 mg/kg,未超标。因此,与自然发酵相比,乳酸菌强化发酵明显减少亚硝酸盐含量,缩短了红酸汤发酵周期,但自然发酵红酸汤品质更好。     

外源碳源对葡萄酵素微生物生长代谢 及生物活性的调节作用

本文以红糖、白糖与蜂蜜为外源碳源添加物,探讨葡萄酵素发酵过程中酵母菌和乳酸菌生长代谢及超氧化物歧化酶(Superoxide dismutase,SOD)和抗氧化活性的差异。采用平板菌落计数法测定微生物菌数;高效液相色谱法(High Performance Liquid Chromatography,HPLC)分析有机酸含量;邻苯三酚法检测SOD酶活;1,1-二苯基-2-三硝基苯肼(1,1-Diphenyl-2-picrylhydrazyl,DPPH)自由基、2,2'-联氮双(3-乙基苯并噻唑啉-6-磺酸)(2,2'-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid),ABTS⁺)自由基和羟基(Hydroxyl radical,OH)自由基清除实验比较抗氧化能力。结果表明,添加碳源减缓酵母生长及乙醇代谢,促进乳酸菌生长及进行产酸活动。其中,红糖、白糖对酵母菌生长及乙醇代谢抑制能力较强,发酵后乙醇浓度各为2.07%vol、1.62%vol,显著低于蜂蜜组(P<0.05);红糖促进乳酸菌生长并提高总酸含量及SOD酶活,发酵后pH由4.16降至3.83,SOD酶活由71.68 U/mL增至91.05 U/mL。与白糖、蜂蜜及对照组相比,添加红糖发酵显著提高葡萄酵素的DPPH、ABTS⁺、OH自由基清除率(P<0.05)。原葡萄液含有大量酒石酸、苹果酸,添加所有碳源发酵显著增加葡萄酵素中苹果酸、及乳酸、乙酸、柠檬酸等含量(P<0.05),但酒石酸含量在添加红糖条件下显著减少(P<0.05),而添加白糖、蜂蜜对酒石酸含量影响不大。因此,添加红糖有利于改善葡萄酵素微生物的生长代谢及提高产品的营养价值。     

Dynamics of the microbial community responsible for traditional sour cassava starch fermentation studied by denaturing gradient gel electrophoresis and quantitative rRNA hybridization

The microbial community developing during the spontaneous fermentation of sour cassava starch was investigated by cultivation-independent methods. Denaturing gradient gel electrophoresis (DGGE) of partially amplified 16S rDNA followed by sequencing of the most intense bands showed that the dominant organisms were all lactic acid bacteria (LAB), mainly close relatives of Bifidobacterium minimum, Lactococcus lactis, Streptococcus sp., Enterococcus saccharolyticus and Lactobacillus plantarum., Close relatives of Lb. panis, Leuconostoc mesenteroides and Ln. citreum were also found. A complementary analysis using hybridization of 16S rRNA with phylogenetic probes was necessary to detect the presence of the recently discovered species Lb. manihotivorans. Although it represented up to 13% of the total lactic acid bacteria of sour cassava starch, this species could not be detected by DGGE as the PCR product migrated to the same position as Lc. lactis. In addition, it was shown that a strong pH decrease in the time course of fermentation was most probably responsible for the competitive selection of acid-resistant LAB vs. both homo and heterofermentative acid-sensitive LAB.     

Microbiological and physicochemical characterisation of green table olives of Halkidiki and Conservolea varieties processed by the Spanish method on industrial scale

The microbiological and physicochemical changes of industrially fermented Halkidiki and Conservolea green table olives were determined. Samples were analysed to monitor the population of lactic acid bacteria (LAB), yeasts and Enterobacteriaceae, together with changes in pH, acidity, salinity, colour, lactic acid, acetic acid and ethanol. LAB and yeast species diversity was evaluated at the beginning (1 day), middle (75 days) and final (135 days) stages of fermentation by RAPD-PCR genomic fingerprinting. Results revealed vigorous lactic acid processes as indicated by the dominance of LAB over yeasts. No Enterobacteriaceae could be detected after 30 days. Lactiplantibacillus plantarum (formerly Lactobacillus plantarum) dominated in the beginning of fermentation in both varieties. In the end, Lactiplantibacillus pentosus (formerly Lactobacillus pentosus) and Pediococcus ethanolidurans prevailed in Halkidiki and Conservolea varieties, respectively. As for yeasts, Kluyveromyces lactis/marxianus and Pichia manshurica prevailed at the onset of fermentation in Halkidiki and Conservolea varieties, whereas in the end Pichia membranifaciens dominated in both varieties.     

Tarhana as a traditional Turkish fermented cereal food. Its recipe, production and composition

As a fermented product tarhana is the dry form of yogurt-cereal mixture and represents an important part of the diets of many people in Turkey. It is prepared by mixing wheat flour, yogurt, yeast and a variety of cooked vegetables (tomatoes, onions, green pepper etc.), salt, and spices (mint, paprika) followed by fermentation for one to seven days. Generally one part yogurt is mixed with two parts of wheat flour (w/w). In commercial production there are two methods for tarhana making. First method is called straight method and ingredients in the recipe is mixed and kneaded, fermented, dried and finally sieved. Second method is called sour dough method that contains three steps, each one has a different recipe. Throughout fermentation lactic acid bacteria and yeast give the characteristic taste and flavour of tarhana by producing lactic acid, ethanol, carbondioxide and some other organic compounds. Organic acids composed in fermentation period lower the pH (3.4-4.2), and low moisture content (6-10%) is a poor medium for pathogens and spoilage organisms. The nutrient content of tarhana depends upon yogurt and flour ratios as well as some other ingredients, and it is also considered to be a useful high-protein dietary supplement with average 15% protein content. Addition of set yogurt due to high dry matter content and baker's yeast increase protein content and enhances it's amino acid composition.     

Chemical studies with silage microorganisms in artificial media and sterile herbages

The fermentation changes in artificial media and in sterile herbages inoculated with strains of yeast, hetero-and homo-fermentative lactic acid bacteria isolated from silage were studied in two experiments. In the first, using malt extract broth, four treatments were employed: aerobic, high level of glucose; aerobic, low level of glucose; anaerobic, high level of glucose; anaerobic, low level of glucose. The pH decreased in all cultures. There were no appreciable differences in pH among the cultures of the heterofermentative lactic acid bacterium, but in the cultures of the homofermentative organism there was a greater decrease in pH aerobically than anaerobically. The lowest pH values in the yeast cultures, occurred in the high glucose treatments both aerobically and anaerobically. In all the yeast cultures, acetic, propionic, butyric, isobutyric and isovaleric acids were present. The heterofermentative lactic acid bacterium produced only acetic acid, the greatest amount occurring under aerobic conditions. Ethanol was detected in the culture of the heterofermentative lactic acid bacterium but not in that of the homofermentative organism. Relatively large amounts of i-pentanol were found in the aerobic yeast cultures. In the second experiment, inoculated grass was ensiled with similar treatments to those used in the first experiment. Two incubation periods, 3d and 10d were used. The pH values in all treatments were high (> pH 5.0). In aerobic conditions, the lactic acid contents after the 10d incubation were higher than after 3d. The losses of water soluble carbohydrates during fermentation were greater in the aerobic than anaerobic treatments. Acetic acid was present in all cultures but the higher volatile fatty acids (C₃-iC₅) were only detected in appreciable amounts in the aerobically treated herbages after 10d. In addition to ethanol, n-propanol and i-butanol were also present in all the 10d anaerobic treatments. i-Pentanol occurred in both the aerobic and anaerobic treated herbages.     

Effect of accelerated natural lactic fermentation of infant food ingredients on some pathogenic microorganisms

Accelerated natural lactic fermentation in mixtures of water and ground sorghum, millet and pigeon pea was obtained by gradual selection of lactic acid bacteria, through inoculum recycling. Weaning food prepared from ingredients fermented this way, contained approx. 0.7% lactic and 0.05% acetic acids and had a pH of about 3.8. In porridges, a pH of less than or equal to 4.0 was required to cause death of Salmonella typhimurium and Staphylococcus aureus. Several intestinal pathogenic bacteria were inoculated into sour porridge. The most resistant Salmonella sp. died at a rate of 1.2 log cycle/h; the most resistant Shigella sp. at 0.9 log cycle/h; and the most resistant Escherichia coli strain at 0.6 log cycle/h. A yeast, Candida albicans, could grow well in the sour product, whereas a bacteriophage (MS-2) was inactivated at a rate of 0.1 log cycle/h. In the acid-sensitive bacterial cultures, no gradual adaptation to acid environments could be observed. The survival studies were carried out at 30 degrees C.     

溶氧(DO)对混菌发酵生产Nisin影响的研究

Nisin(乳酸链球菌素)是由乳酸菌产生的一种抑菌活性肽。其在生产过程中需要控制发酵体系的pH,以避免由于乳酸菌代谢积累的乳酸导致pH迅速降低抑制菌体生长。本文采用以能够代谢消耗乳酸的酵母菌为辅助发酵菌的新型混菌发酵方法,在小试条件下研究了在混菌发酵过程中对菌体特别是酵母菌代谢有显著影响的因素溶氧(DO)对混菌发酵生产过程中发酵体系的pH与乳酸含量的变化nisin产量的影响。结果表明,不同水平溶氧对混菌发酵体系中两种菌的生长及代谢产物生成有一定影响。DO 10%与DO 30%时的nisin效价的水平波动范围为24.35%;在分阶段控制发酵体系溶氧水平的条件下获得了较高的nisin效价,效价水平比恒定溶氧提高45.38%。说明分阶段控制发酵体系溶氧水平是较为理想的溶氧控制策略。     

酸肉发酵中蛋白质降解及影响因素的研究

目的：了解蛋白质降解与酸肉品质间的关系以指导合理消费。方法：分析酸肉在发酵过程中主要环境因素、乳酸菌数及蛋白质降解产物的变化。结果：总蛋白质、肌原纤维蛋白和肌浆蛋白随发酵时间延长逐渐下降,非蛋白氮逐渐增加;蛋白质降解中间产物多肽氮及氨态氮呈先升后降趋势,发酵60d时挥发性盐基氮为19.56g/100g;氨基酸组成分析显示,发酵可提高蛋白质氨基酸分;相关性分析显示总蛋白质、肌原纤维蛋白和肌浆蛋白含量变化与乳酸菌数和氯化钠含量变化呈显著负相关,而与水分活度和pH值呈显著正相关;非蛋白氮与乳酸菌数和氯化钠含量显著正相关,而与水分活度和pH值显著负相关。结论：发酵可提高酸肉蛋白质营养价值,综合安全及风味因素,建议酸肉发酵时间在20～40d为宜。     

白首乌酵素发酵工艺的优化

为了开发一款白首乌酵素产品,本文以白首乌为原料,采用乳酸菌和酵母菌复合发酵制备酵素,对乳酸菌和酵母菌的单因素实验条件进行优化,并确定两菌种复合发酵时的接种顺序和发酵时间。结果表明:采用先接种酵母菌后接种乳酸菌发酵的方式最优,该接种顺序下的最佳复合发酵工艺条件为:梅山酵母菌(S4)在初始pH5.0、装瓶量30 mL(250 mL)、蛋白胨添加量1.2%、接种量0.2%、发酵温度30 ℃的条件下,发酵24 h,酵母菌发酵结束后,接种3%的复合乳酸菌(嗜酸乳杆菌(LA)和植物乳杆菌(LP)以1:1混合(v:v)),37 ℃静置发酵36 h,然后在4~8 ℃下后发酵12 h,使发酵液产香。在此条件下,白首乌酵素中乳酸菌活菌为2.61×10⁸ CFU/mL,酵母菌活菌数为7.9×10⁷ CFU/mL,超氧化物歧化酶(superoxide dismutase,SOD)活力为4.23×10⁴ U/L。采用发酵的方法制备白首乌产品,为白首乌资源的开发利用提供了新的思路,对促进白首乌产业的发展具有重要意义。