Production of conjugated linoleic acid by lactic acid bacteria in milk without any additional substrate

This study was undertaken to evaluate the production and isomeric distribution of conjugated linoleic acid (CLA) in milk fermented with lactic acid bacteria (LAB. A total of 155 cultures of LAB were analysed. Control milk samples had an average CLA content of 0.41 g/100 g of fatty acids (FA), while the lactic cultures produced CLA in the range of 0.43–1.12 g. No major changes in free fatty acids profiles were observed in milk samples fermented with CLA‐producing LAB. The present study demonstrated that LAB can increase levels of CLA in nonsupplemented milk fermented for a short period of 4 h.     

植物乳杆菌转化合成共轭亚油酸的培养基条件优化

生物法转化合成共轭亚油酸,产物中异构体组成单一,具有很好的应用前景。通过一株植物乳杆菌(Lactobacillus plantarum)合成共轭亚油酸的培养基成分进行优化,确定最优的培养基组成:葡萄糖20g/L,酵母浸出物40g/L,硫酸镁0.5g/L,硫酸锰0.5g/L,乙酸钠2g/L,磷酸氢二钾1g/L。优化后,共轭亚油酸产量达到0.259g/L,相比优化前(0.0455g/L)有了较大的提升。      

乳酸菌洗涤菌体合成共轭亚油酸的研究

为了解瑞士乳杆菌L7(Lactobacillus helveticusL7)洗涤菌体生物合成共轭亚油酸(CLA)的能力,通过单因素和正交优化研究,确定了瑞士乳杆菌L7洗涤菌体合成c9,t11-CLA的最适条件:0.05mol/LPBS缓冲液(pH5.8)、1.00mg/mL亚油酸、23℃反应12h。在最适转化条件下,c9,t11-CLA产量达到0.54mg/mL。结果表明,洗涤菌体合成CLA的产量和分批发酵相近,可以继续进行瑞士乳杆菌L7固定化细胞发酵生产CLA的研究。     

植物乳杆菌ZS2058生物转化共轭亚油酸反应条件的优化研究

对本实验室从泡菜中筛选到的植物乳杆菌ZS2058完整细胞生物转化共轭亚油酸的反应条件进行了系统研究。在1mL磷酸钾缓冲液反应体系中,通过单因素实验和响应面分析,确定最合适的反应条件为:亚油酸底物浓度为0.8mg/mL,细胞浓度为4×1010cfu/mL,反应温度为37℃,缓冲液pH为6.7。在此反应条件下,cis9,trans11-CLA的浓度为374.24μg/mL,转化率高达46.78%,这对于实现共轭亚油酸的高效生产和研究其生理功能具有重要的现实意义和理论价值。      

Influence of sodium glycocholate on production of conjugated linoleic acid by cells of Lactobacillus reuteri ATCC 55739

ABSTRACT:  The possible influence of a bile salt on production of conjugated linoleic acid (CLA) by Lactobacillus reuteri ATCC 55739 was evaluated. Cells of the lactobacilli grown in MRS broth with and without linoleic acid (LA, 0.2%) were harvested and washed. The washed cells were added to buffer containing 0.2% LA and incubated 18 h at 37 °C. The cells, which had been grown without LA, transformed LA into CLA (mainly c9t11-C18:2) better than did those cells grown with it. When sodium glycocholate (0.3%) was added to the washed cell suspensions, about the same level of CLA was formed as in its absence regardless of whether or not the cells had been grown in broth supplemented with free LA. Thus, glycocholate that occurs in humans did not influence production of CLA by resting cells of the lactobacilli.     

Production of conjugated linoleic acid by Propionibacterium freudenreichii

Two strains each of Propionibacterium freudenreichii ssp. shermanii and P. freudenreichii ssp. freudenreichii were tested for their ability to produce conjugated linoleic acid (CLA) in sodium lactate medium (SLM), De Man-Rogosa-Sharpe (MRS) medium and skim-milk. Data showed that both strains were able to produce CLA in three media supplemented with different concentrations of sunflower oil. Maximum production of CLA (78.8 μg/ml) was observed after 36 h of incubation in MRS containing 12 mg/ml of sunflower oil by P. freudenreichii ssp. shermanii. Moreover, the growths of both strains were inhibited by sunflower oil and a positive relationship between CLA production and ability to tolerate sunflower oil was observed. At the same time, it was also observed that the inhibitory effects on P. freudenreichii ssp. shermanii and P. freudenreichii ssp. freudenreichii in three media follow the order SLM > skim-milk > MRS and SLM > MRS > skim-milk, respectively. Micro aerobic conditions were in favour of increasing the amounts of CLA. The amounts of CLA increased from 0 to 36 h under micro aerobic conditions and no significant (p > 0.05) increases in total CLA levels were observed after 80 h of incubation. Results showed that P. freudenreichii may have potential for producing CLA.     

牛奶中共轭亚油酸含量的紫外分光光度测定方法

研究了牛乳中共轭亚油酸含量的紫外分光光度测定方法。利用紫外分光光度计,先建立标准曲线与回归公式,再将牛奶样品提取、甲酯化后,进行紫外检测。结果表明,共轭亚油酸甲酯的最大吸收波长是234nm;共轭亚油酸浓度的有效测定范围是0.00856-0.0428mg/mL;吸光度与共轭亚油酸浓度间的回归公式为Y=52.602X 0.0917(R2=0.9862);得出共轭亚油酸浓度与吸光度之间的经验公式为:共轭亚油酸浓度C(mg/mL)=0.019×吸光度(ABS);并且推出了利用紫外分光光度法测定牛奶中共轭亚油酸含量的计算公式。同时,检验了几种市售牛奶中共轭亚油酸,其平均含量为1.64mg/g。     

植物乳杆菌ZS2058转化亚油酸为共轭亚油酸条件的初步研究

研究了植物乳杆菌ZS2058转化亚油酸为共轭亚油酸的条件,在培养基中添加10g/L吐温-80制成LA-吐温-80胶束溶液,可以提高CLA的转化率,最佳转化温度为35℃,添加1mg/mL的LA有较高的转化率和CLA产量。在MRS、SKM、KPB三种转化体系中SKM的转化率较高。在含LA的培养基中预培养12h,可以提高CLA的转化率。最佳转化接种量在1%～2%之间。      

嗜酸乳杆菌转化菜籽油生成共轭亚油酸条件的优化

文中对嗜酸乳杆菌静息细胞转化菜籽油生成CLA工艺进行了优化研究,通过单因素试验,确定了最适转化液介质、介质浓度、pH、温度、转化时间和细胞浓度,同时,研究了不同金属离子对亚油酸异构酶活性的影响。在该基础上,利用Box-Benhnken中心组合设计和响应面法对转化时间、pH、温度、细胞浓度进行了工艺优化分析,确定了在菜籽油浓度为7.5 mg/mL、脂肪酶的加入量为2 mg/30 mL时,以0.1 mol/L柠檬酸-柠檬酸钠缓冲液为转化液介质条件下,最优转化工艺条件是时间25 h,pH6.5,温度36℃,细胞浓度40 mg/mL,经优化后CLA的生成量可以达到230.12±7.52μg/mL。     

气相色谱法测定牛奶中c9,t11-共轭亚油酸的含量

为建立简单高效的分析方法测定牛奶中c9,t11-共轭亚油酸的含量,应用GC-FID法,采用CP-Sil88(60m×0.25mmi.d×0.20μm)气相色谱柱,以程序升温方法,应用带校正因子的面积归一法进行定量分析。结果表明,c9,t11-共轭亚油酸得到很好的分离,平均回收率为97.7%,RSD为1.08%。     

Volatile compounds of fresh and dry-cured loin as affected by dietary conjugated linoleic acid and monounsaturated fatty acids

Three levels (0%, 1% and 2%) of an enriched conjugated linoleic acid oil (CLA) were combined with two levels of monounsaturated fatty acids (MUFA) (low (19%) average and high (39%) average) for pig feeding. The profile of volatile compounds of fresh and dry-cured loin as affected by dietary CLA, MUFA and CLA × MUFA interaction was studied by headspace-solid phase microextraction coupled to gas chromatography–mass spectrometry. A total of 27 and 69 compounds were identified in fresh loin and dry-cured loin, respectively. Identified compounds were alcohols, aldehydes, aliphatic hydrocarbons, aromatic hydrocarbons, esters, furans, ketones, nitrogen compounds, sulphur compounds and terpenes. No qualitative differences in volatile compounds caused by the assayed treatments were found neither in fresh loin nor in dry-cured loin. Dietary CLA, MUFA and their interaction did not affect the level of most detected volatiles. Nevertheless, in fresh loin, the level of heptanal significantly increased due to dietary CLA (p = 0.001) and the area units (AU) for ethyl benzene and 1,3-dimethyl benzene increased at 1% CLA (p < 0.05). In the case of dry-cured loin, the AU of heptanal, nonanal and 2-nonenal contents increased due to dietary CLA (p = 0.016, 0.024 and 0.019, respectively).     

Phytate dephosphorylation by Lactobacillus pentosus CFR3

Studies on phytate‐degrading enzymes from lactobacilli are scarce, despite its potential in improving the nutritional quality of plant‐based foods. Therefore, the current investigation deals with the phytate‐degrading enzyme produced by a native Lactobacillus pentosus strain. Phytase activity was highest towards the end of the exponential phase. Activity increased in the presence of maltose (381.1%) compared with glucose. The presence of phytate in the media stimulated the enzyme production. The enzyme of interest was a 70 kDa protein with a pH and temperature optima of 5.0 and 55–60 °C, respectively. It retained 46% of activity after exposure to 70 °C for 20 min and also showed broad substrate specificity. It was completely inhibited by Hg²⁺, Fe²⁺ and PMSF while being activated by Co²⁺. This report is the first to show dephytinisation of autoclaved finger millet flour either by fermentation with L. pentosus or by treatment with the corresponding cell‐free extract.     

Conjugated linoleic acid content in fermented goat milk as affected by the starter culture and the presence of free linoleic acid

The effect of yoghurt starter cultures and probiotic Lactobacillus casei on the formation of conjugated linoleic acid (CLA) and microbial populations of fermented goat milk was investigated during 35 days of cold storage. The addition of hydrolysed sunflower oil as a source of free linoleic acid was investigated. The fermentation process enhanced the content of the cis9, trans11‐CLA isomer in milk, whereas the trans10, cis12‐CLA isomer was not detected in goat milk or control fermented milks. The use of both starters generated trans10, cis12‐CLA only when hydrolysed sunflower oil was supplemented. Populations of streptococci and lactobacilli were affected by the presence of hydrolysed sunflower oil.     

Production of bioactive substances by intestinal bacteria as a basis for explaining probiotic mechanisms: bacteriocins and conjugated linoleic acid

The mechanisms by which intestinal bacteria achieve their associated health benefits can be complex and multifaceted. In this respect, the diverse microbial composition of the human gastrointestinal tract (GIT) provides an almost unlimited potential source of bioactive substances (pharmabiotics) which can directly or indirectly affect human health. Bacteriocins and fatty acids are just two examples of pharmabiotic substances which may contribute to probiotic functionality within the mammalian GIT. Bacteriocin production is believed to confer producing strains with a competitive advantage within complex microbial environments as a consequence of their associated antimicrobial activity. This has the potential to enable the establishment and prevalence of producing strains as well as directly inhibiting pathogens within the GIT. Consequently, these antimicrobial peptides and the associated intestinal producing strains may be exploited to beneficially influence microbial populations. Intestinal bacteria are also known to produce a diverse array of health-promoting fatty acids. Indeed, certain strains of intestinal bifidobacteria have been shown to produce conjugated linoleic acid (CLA), a fatty acid which has been associated with a variety of systemic health-promoting effects. Recently, the ability to modulate the fatty acid composition of the liver and adipose tissue of the host upon oral administration of CLA-producing bifidobacteria and lactobacilli was demonstrated in a murine model. Importantly, this implies a potential therapeutic role for probiotics in the treatment of certain metabolic and immunoinflammatory disorders. Such examples serve to highlight the potential contribution of pharmabiotic production to probiotic functionality in relation to human health maintenance.     

Amounts of conjugated linoleic acid (CLA) in German foods and evaluation of daily intake

 The quantities of the biologically active isomer of conjugated linoleic acid (CLA) – C18:2 c9t11 – in 139 German foods were analysed by capillary gas chromatography (results are given as a % of all identified fatty acid methyl esters). The CLA content ranged from 0.40% (Gouda) to 1.70% (Jurassic cheese, Old Emmentaler) in dairy products, from 0.11% (rabbit) to 1.20% (lamb) in meat, and from 0.01% (pike-perch) to 0.09% (carp) in fish. CLA could be detected in neither vegetable fats or oils nor in margarines (CLA <0.01%). Crisps, chocolates, cakes and pastries, and other foods have only a negligible CLA content. The average estimated CLA intake in Germany was calculated to be 0.35g CLA/day for women and 0.43g CLA/day for men. Received: 22 July 1997     

Amounts of conjugated linoleic acid (CLA) in German foods and evaluation of daily intake

 The quantities of the biologically active isomer of conjugated linoleic acid (CLA) – C18:2 c9t11 – in 139 German foods were analysed by capillary gas chromatography (results are given as a % of all identified fatty acid methyl esters). The CLA content ranged from 0.40% (Gouda) to 1.70% (Jurassic cheese, Old Emmentaler) in dairy products, from 0.11% (rabbit) to 1.20% (lamb) in meat, and from 0.01% (pike-perch) to 0.09% (carp) in fish. CLA could be detected in neither vegetable fats or oils nor in margarines (CLA <0.01%). Crisps, chocolates, cakes and pastries, and other foods have only a negligible CLA content. The average estimated CLA intake in Germany was calculated to be 0.35g CLA/day for women and 0.43g CLA/day for men. Received: 22 July 1997     

Effects of dietary replacement of fish oil by conjugated linoleic acid on some meat quality traits of Pacific white shrimp Litopenaeus vannamei

This experiment was conducted to evaluate the effects of fish oil replacement by conjugated linoleic acid (CLA) on body proximate analysis and thiobarbituric acid-reactive substances, fat content, shear force and fatty acid composition in musculature of Pacific white shrimp (Litopenaeus vannamei). Graded levels of CLA (0%, 0.5%, 1% and 2%) were added to the basic diet of shrimp at the expense of fish oil. Results showed that fat content (p = 0.036) and shear force (p = 0.001) in shrimp musculature were enhanced with increasing dietary CLA inclusion. Fish oil replacement by CLA significantly promoted the incorporation of cis-9, trans-11 CLA (p = 0.0001) and trans-10, cis-12 CLA (p < 0.0001) into shrimp musculature; moreover, the polyunsaturated fatty acid was elevated (p = 0.020) and monounsaturated fatty acid was reduced by CLA inclusion (p = 0.024). It was concluded that replacement of fish oil by CLA could improve some meat quality traits of shrimp and 1% CLA was an appropriate amount.     

Seasonal variation in conjugated linoleic acid and vaccenic acid in milk fat of sheep and its transfer to cheese and ricotta

The seasonal variation in conjugated linoleic acid (CLA) and vaccenic acid (VA) concentrations in sheep dairy products and the extent of their transfer from milk fat to cheese and ricotta fat were investigated. Samples were collected from 2 sheep milk processing plants in North Sardinia (Italy) every 2 wk from March through June. Concentrations of fatty acids (FA) in fresh cheese and ricotta fat were primarily dependent on the fatty acid content of the unprocessed raw milk. The content of c9,t11-CLA averaged 1.73, 1.69, and 1.75 mg/100 mg of FA methyl esters (FAME), respectively, for milk, cheese, and ricotta, and differed significantly between cheese and ricotta. The content of VA averaged 3.40, 3.33, and 3.43 mg/100 mg of FAME, respectively for milk, cheese, and ricotta. The FA composition of dairy products was markedly affected by period of sampling: the mean c9,t11-CLA and VA concentration decreased from March (2.20 and 4.52 mg/100 mg of FAME) to June (1.14 and 1.76 mg/100 mg of FAME) in all dairy products. No differences in c9,t11-CLA and VA concentration of dairy products were observed between the 2 dairy companies obtaining milk from the same geographical origin. The seasonal changes in CLA and VA in milk fat were probably related to changes in pasture quality.     

Volatile organic compounds associated with milk spoilage by psychrotrophic bacteria

The volatile organic compounds of milk contaminated with psychrotrophic bacteria were studied by HS‐SPME and GC/MS. Pseudomonas fluorescens PS14, Pseudomonas fragi PS55, Pseudomonas mosselii PS39, Pseudomonas rhodesiae PS62 and Serratia marcescens S92 were inoculated in sterilised milk (2.5% fat) stored at either 5 °C or 10 °C. A total of 47 volatile organic compounds (VOCs) belonging to seven chemical groups were identified in the spoiled milk. Volatile organic compound patterns peculiar to the inoculate bacterial strains were highlighted. 3‐Methylbutan‐1‐ol, 2 methylpropan‐1‐ol, 3‐hydroxybutan‐2‐one, butan‐2,3‐dione, butanoic and hexanoic acids were revealed as potential chemical spoilage indexes of milk spoilage due to the activity of the five psychrotrophic strains studied.