Linoleic acid Isomerase Activity in Enzyme Extracts from Lactobacillus Acidophilus and Propionibacterium Freudenreichii ssp. Shermanii

Enzyme extracts from Lactobacillus acidophilus (CCRC14079) and Propionibacterium freudenreichii ssp. shermanii (CCRC11076) were reacted with linoleic acid at 50 °C for 10 min at pH 5, 6, 7, and 8, and the levels of conjugated linoleic acid (CLA) formation were determined by high performance liquid chromatography. CLA formations were observed in all the reactions catalyzed by the retentates using ultrafiltration membranes with 100 kDa nominal molecular weight cutoff, indicating the presence of the activity of linoleic acid isomerase with nominal molecular weight higher than 100 kDa in the retentates from two cultures. More CLA was formed at pH 5 of L. acidophilus treatment and t10c12, c11t13, and c9t11 CLA were 3 major CLA isomers produced. Results demonstrate a potential for CLA production through linoleic acid isomerase.     

Concentration‐dependent antioxidant activities of conjugated linoleic acid and α‐tocopherol in corn oil

BACKGROUND: Antioxidants prevent rancidity (lipid peroxidation) and natural antioxidants, e.g., α‐tocopherol, likely provide additional value to oil‐based food products because of their health benefits. Conjugated linoleic acid (CLA) has potential health benefits and may exhibit antioxidant properties. The main aim of this study was to compare the antioxidant efficacy of α‐tocopherol, trans‐10, cis‐12‐CLA and cis‐9, trans‐11‐CLA (in graded concentrations) added to antioxidant‐stripped corn oil. RESULTS: As compared to α‐tocopherol, both CLA isomers displayed significant inhibition of corn oil lipid peroxidation induced by copper. Inhibition of thiobarbituric acid reactive substances (TBARS) were CLA concentration dependent for both isomers but with significant inhibition occurring at 0.1 and 1 ppm of CLA isomers or α‐tocopherol, respectively (P < 0.05). Graded concentrations of α‐tocopherol, and for both CLA isomers and time, had significant effects on TBARS formation (P < 0.0001). There were significant effects in interactions between graded concentrations and time for both CLA isomers (P < 0.0001) but not for α‐tocopherol (P > 0.05). CONCLUSION: CLA compounds could serve as useful food antioxidants and provide additional value because of their potential bioactivity in disease prevention. Copyright © 2007 Society of Chemical Industry     

Advances in Research on cis-9, trans-11 Conjugated Linoleic Acid: A Major Functional Conjugated Linoleic Acid Isomer

Conjugated linoleic acid (CLA) consists of a group of positional and geometric conjugated isomers of linoleic acid. Since the identification of CLA as a factor that can inhibit mutagenesis and carcinogenesis, thousands of studies have been conducted in the last several decades. Among the many isomers discovered, cis-9, trans-11 CLA is the most intensively studied because of its multiple, isomer-specific effects in humans and animals. This paper provides an overview of the available data on cis-9, trans-11 CLA, including its isomer-specific effects, biosynthesis, in vivo/in vitro research models, quantification, and the factors influencing its content in ruminant products.     

Effect of incubation conditions and possible intestinal nutrients on cis-9, trans-11 conjugated linoleic acid production by Lactobacillus acidophilus F0221

The purpose of this study was to optimise the incubation conditions for cis-9, trans-11 conjugated linoleic acid (c9, t11 CLA) production by Lactobacillus acidophilus F0221 and evaluate the effect of possible intestinal nutrients on c9, t11 CLA production. Growth in Mann Rogosa Sharp broth supplemented with cysteine and containing 0.5 g L⁻¹ linoleic acid at pH 6.5 with anaerobic incubation at 37 °C for 40 h were found to be the optimal conditions for CLA production. Galactosaccharide, arabinogalactan, galactose and glucose had a higher promoting effect on CLA production (96.19–123.89 μg mL⁻¹) than other carbohydrates. Acetic acid had a higher promoting effect on CLA production (130.98 μg mL⁻¹) than other short chain fatty acids. These results provide detailed parameters for the production of c9, t11 CLA by L. acidophilus F0221 and are valuable for further understanding of c9, t11 CLA formation in the human intestine by this strain.     

Effect of probiotic co-cultures on physico-chemical and biochemical properties of small ruminants’ fermented milk

Small ruminants' fermented probiotic milk is an alternative to fermented cows' milk, especially because of the monounsaturated/polyunsaturated fatty acid profiles. The technological and biochemical potential of Bifidobacterium and Lactobacillus co-cultures, with or without inulin, on goats' and ewes' milk was assessed. Microbial stability, lactose consumption, organic acid production, proteolytic parameters and conjugated linoleic acid (CLA) production in situ, were followed in ewes' and goats’ fermented milk (EFM and GFM, respectively) over 21 days at 4 °C; technological feasibility for probiotic fermented milk production was shown. In EFM, all co-cultures presented high viable cell numbers (>7.0 log cfu mL⁻¹) throughout storage, presenting faster acidification capacities and higher CLA isomer levels than in GFM. Inulin had no impact on probiotic growth, yet contributed to storage stability. CLA isomers and proteolysis indices were co-culture dependent traits: for example, co-culture of Bifidobacterium animalis B94 with Lactobacillus acidophilus L10 registered the best CLA-production in GFM.     

Effect of Conjugated Linoleic Acids in Pig Nutrition on Quality of Meat

Relationships among conjugated linoleic acid (CLA) in pig nutrition and carcass quality parameters (hot carcass weight, carcass yield and meatiness) and meat quality parameters (initial and pH value after 24, 48 and 72 h, temperature, drip loss, sensory color and marbling) were determined in pigs (crossbreeds Yorkshire x Landrace). Commercial CLA preparation containing 60% CLA isomers was included in the diet. No significant differences in performance parameters were found between pigs fed with CLA and control group during 60 days period. CLA supplementation in feed significantly increased SFA and decreased MUFA and PUFA fraction in pig muscles.     

Conjugated linoleic acid production by cells and enzyme extract of Lactobacillus delbrueckii ssp. bulgaricus with additions of different fatty acids

The objective of the study was to determine the effect of fatty acid additions to the cells and enzyme extract of Lactobacillus delbrueckii ssp. bulgaricus (CCRC14009) on CLA production. Washed cells of L. delbrueckii ssp. bulgaricus, obtained by cultivation in a MRS broth, were mixed with BSA and each of the three fatty acids: linoleic, oleic, and linolenic acids in sodium phosphate buffer at pH 6.5. After incubation at 37 °C for 108 h, CLA concentration was analyzed by HPLC. Enzyme extract from the culture was also reacted with each fatty acid at 50 °C for 10 min at pH 5 to test for CLA production. Results showed that linoleic acid addition to the culture improved CLA production, indicating the presence of linoleic acid isomerase activity in the culture. The crude enzyme extract from the culture was observed to be capable of oleic and linolenic acid conversions into CLA, demonstrating the possible presence of desaturase activity in the enzyme extract.     

Grass-based ruminant production methods and human bioconversion of vaccenic acid with estimations of maximal dietary intake of conjugated linoleic acids

Reports on human conjugated linoleic acids (CLAs) intake vary, and usually don't consider food production methods and vaccenic acid (VA)-induced CLA biosynthesis in humans. The aim of this review was to estimate the achievable CLA intake considering both food sources produced by feeding patterns natural to animals and human VA bioconversion. Exogenous CLA supply from milk, cheese, lamb, and beef from grass-based ruminant production methods, was calculated to be about double that of estimates based on modern production methods (respectively, from the four sources: 1.25, 1.50, 1.44, and 0.69% of total fatty acids). Using available human consumption data this resulted in an estimated achievable CLA intake (including VA bioconversion) of between 711 and 1107 mg d^?1. Intake of products from natural grass-fed ruminants leads to a several-fold higher CLA food content and, along with consideration of endogenously formed CLA from VA, results in a substantially higher CLA availability than previously estimated.     

Conjugated linoleic acid production by immobilized cells of Lactobacillus delbrueckii ssp. bulgaricus and Lactobacillus acidophilus

Lactobacillus delbrueckii ssp. bulgaricus (CCRC14009) and L. acidophilus (CCRC14079), immobilized with chitosan and polyacrylamide, were tested for CLA production. A 10-ml aliquot of L. delbrueckii ssp. bulgaricus cell suspension (3.59 × 10⁷ CFU/ml) was adsorbed to 0.5 g chitosan and polyacrylamide, mixed with 0.2 ml linoleic acid (0.9 g/ml), and incubated at 37 °C for 24 h at pH 5, 6, 7, and 8 for CLA production. CLA levels, produced by immobilized cells of L. delbrueckii ssp. bulgaricus and L. acidophilus with increasing cell counts to 1.08 and 1.28 × 10¹⁰ CFU/ml, respectively, at optimal reaction pHs were evaluated. More CLA was formed at pH 8 of chitosan and pH 7 of polyacrylamide-immobilized L. delbrueckii ssp. bulgaricus cell treatments. Increase in cell count resulted in higher CLA production. The adsorption of L. delbrueckii ssp. bulgaricus cells onto polyacrylamide at pH 7 showed significant improvement in total CLA level. Results demonstrated a potential for enhancing CLA production through immobilization.     

The triacylglycerol preparation of conjugated linoleic acid reduces lipid oxidation in irradiated, cooked ground beef patties

It is proposed that conjugated linoleic acid (CLA) would depress the lipid oxidation caused by irradiation of cooked, aerobically stored ground beef patties. The free fatty acid (FFA–CLA) and triacylglycerol (TAG–CLA) preparations of CLA were added at 0%, 1%, 2%, or 4% during the grinding process. Patties were irradiated at 1.5–2.0 kGy and frozen at −20 °C. Subsequently, the patties were tempered to 4 °C, cooked to 70 °C and held at 4 °C for 7 d. Enrichment of ground beef with CLA increased the cis-9,trans-11 and CLA trans-10,cis-12 CLA isomers in ground beef patties, even after cooking. Weight loss (P = 0.03) and percentage fat (P = 0.05) were higher in irradiated beef patties than in control patties. Irradiation decreased the concentration of α-linolenic acid (18:3n − 3) in the ground beef by over 60% (P = 0.07), whereas thiobarbituric acid reactive substances (TBARS) values were higher (P = 0.004) in irradiated beef patties than in control patties. The 1% concentration of added TAG–CLA reduced TBARS in irradiated ground beef patties, whereas 2% and 4% FFA–CLA depressed TBARS (CLA type × percentage interaction P = 0.04). Irradiation increased the cardboard and painty aromatic attributes (P  0.05), and FFA–CLA preparation increased the painty aromatic attribute and afterburn aftertaste, but these effects were not observed with the TAG–CLA preparation (CLA type × treatment interaction P < 0.04). Adding 1% TAG–CLA to ground beef during grinding can reduce lipid oxidation in irradiated, cooked ground beef patties without the negative aftertastes associated with the FFA–CLA preparation.     

共轭亚油酸的生理功能及其合成、纯化研究检测方法

共轭亚油酸是一系列位置和几何异构体的总称，其中的活性异构体具有多种重要的生理功能，近年来受到越来越多的重视。对活性共轭亚油酸异构体的主要生理功能、微生物合成共轭亚油酸的微生物菌种及合成途径中的关键性酶、目前使用的共轭亚油酸的化学合成方法、以及共轭亚油酸的纯化及分析检测方法进行了综述。     

Evidences and perspectives in the utilization of CLNA isomers as bioactive compounds in foods

Conjugated alpha linolenic acid (CLNA) isomers are promising lipids owing to their similarities with conjugated linoleic acid (CLA) but exerting their bioactivity at lower doses; some isomers also belong to omega 3 family. This review aims to summarize the state of the art about the utilization of CLNA as a functional ingredient. Indeed, in vitro and in vivo studies reported that CLNA exerted anticancer, anti-inflammatory, anti-obese, and antioxidant activities. However, CLNA has not been tested in humans. These compounds are naturally present in meat and milk fat from ruminants but the highest concentrations are found in vegetable oils. Their incorporation in foodstuffs is one of the most effective strategies to elaborate CLNA-enriched products together with the microbiological production. Lactobacilli, propionibacteria, and bifidobacteria strains have been assayed to produce CLNA isomers but at the current moment there are not high CLNA concentration products elaborated using these strains. Furthermore, it is known that CLNA isomers are highly prone to oxidation when compared with linoleic acid and CLA, but the possible effects of elaboration and storage on high CLNA productsare unknown.The utilization of CLNA as a functional compound still remains a challenge and requires more research to address all of its technological and bioactivity aspects.     

共轭亚油酸的合成

共轭亚油酸(CLA)是亚油酸(LA)的同分异构体,是一种非常重要的功能性油脂,在医药保健、食品工业、饲料等方面的应用有着广阔的应用前景.从CLA的生产方法着手,重点描述了传统CLA的制备方法-化学异构化法、羟基脂肪酸脱水法,生物合成法和光异构化法.并对这些方法的研究手段、国内外研究水平、存在的问题、解决措施、优缺点和应用前景进行了介绍.尤其是生物合成法,由于其产物选择性高、异构体种类少、培养易于控制等,具有很好的商业应用前景.     

Influence of a dry fractionation of butterfat on the content of fatty acids including conjugated linoleic acids

There is a growing demand among consumers for food products with natural nutritional–physiological advantages over comparable conventional products. As part of an EU project, a process using dry fractionation is evaluated that enables the targeted low-input enrichment of conjugated linoleic acids (CLA) in milk fat. Furthermore, the distribution of CLA isomers in the fat fractions was analysed. In the olein fraction for highland butter a CLA enrichment of 15.3% was obtained. The yield of the CLA rich olein fraction was 44.5% of the total amount of olein and stearin. There were significant increases during the first fractionation step of highland butter for the concentration of the CLA isomer cis-9, trans-11 (P ≤ 0.05) and during the second fractionation step for the concentration of CLA isomers cis-9, trans-11; trans-11, cis-13 (P ≤ 0.05) and trans-7, cis-9 (P ≤ 0.01). Experiments carried out demonstrate that the selected physical separation process enables CLA enrichment but the increase is too minor to achieve any decisive positive impact on human health and therefore too costly as an industrial CLA enrichment process.     

Production and structural analysis of triacylglycerols containing capric acid and conjugated linoleic acid isomers obtained by enzymatic acidolysis

BACKGROUND: Structured lipids containing medium‐chain fatty acids have interesting applications as reduced‐calorie fats; moreover, conjugated linoleic acid (CLA) isomers have shown interesting biological properties. The aim of this study was to synthesize triacylglycerols (TAGs) containing capric acid in the sn‐1‐ and sn‐3‐ positions and CLA isomers in the sn‐2‐ position, using different commercial available lipases. RESULTS: The homogeneous CLA‐TAGs (Tri‐CLA) were chemically synthesized starting from glycerol and CLA isomers, 9‐cis,11‐trans and 10‐trans,12‐cis CLA. The acidolysis reactions of Tri‐CLA with capric acid were carried out at 55 °C for different times in hexane; after 96 h the acidolysis average yield was 65%. The best capric acid incorporation in total TAGs was obtained after 96 h with Lipozyme IM (56.6%). The results of structural analysis carried out on the obtained TAGs showed that both Novozyme 435 and anhydrous Lipozyme IM gave the best incorporation of capric acid in sn‐1(3)‐ positions (61.8%). However, anhydrous Lipozyme IM gave also the highest CLA percent content in sn‐2‐ position (73.2%). CONCLUSION: Anhydrous Lipozyme IM appears to be the more effective enzyme in acidolysis reactions to obtain structured TAGs containing CLA isomers in the central position and capric acid at external positions. Copyright © 2009 Society of Chemical Industry     

In vitro characterisation of probiotic properties of Enterococcus faecium and Enterococcus durans strains isolated from raw milk and traditional dairy products

In this study, some probiotic characteristics such as antimicrobial activity, vancomycin resistance, growth ability at pH, resistance to bile salts, bile salt deconjugation and hydrophobicity of 30 Enterococcus faecium and Enterococcus durans strains isolated and identified from raw milk and various dairy products were investigated. According to the study results, antimicrobial activity profiling, pH and bile salt resistance and bile salt deconjugation ability of Enterococcus strains varied depending on the species and strains and all the strains showed resistance to the tested bile salt concentrations. It was concluded that the E. faecium and E. durans strains tested showed probiotic characteristics and have the potential to be used in food production.     

Production of free conjugated linoleic acid by Lactobacillus acidophilus and Lactobacillus casei of human intestinal origin

A gas chromatographic procedure was used for analysis of conjugated linoleic acid (CLA) isomers cis-9, trans-11-octadecadienoic; trans-10, cis-12 octadecadienoic; and trans-9, trans-11-octadecadienoic (c9t11, t10c12, t9t11) produced by lactobacilli. Four different cultures, two strains each of Lactobacillus acidophilus and Lactobacillus casei were tested for their ability to produce CLA from free linoleic acid in MRS broth supplemented with linoleic acid. Different concentrations of linoleic acid (0, 0.05, 0.1, 0.2 and 0.5 mg/ml) were added to MRS broth, inoculated with the lactobacilli, and incubated at 37 degrees C. Viable counts and amounts of individual isomers of CLA (c9t11, t10c12, t9t11) were measured at 0, 24, 48, and 72 h. All the cultures were able to produce free CLA in media supplemented with linoleic acid. Maximum production of CLA (80.14 to 131.63 microg/ml) was observed at 24 h of incubation in broth containing 0.02% of free linoleic acid. No significant (P > 0.05) increases in total CLA levels were observed after 24 h of incubation. The ability of the cultures to produce CLA in skim milk supplemented with 0.02% free linoleic acid also was studied. In this medium, the total amounts of free CLA after 24 h of incubation ranged from 54.31 to 116.53 microg/ml. The use of lactic acid bacteria able to form free CLA in cultured dairy products may have potential health or nutritional benefits. Free CLA in the products likely would be more readily available for absorption from the digestive tract than if it were incorporated into the cells of the starter culture.     

Inulin and probiotic concentration effects on fatty and linoleic conjugated acids in cream cheeses

The aim of this study was to evaluate the changes in the lipid profile, especially in conjugated linoleic acid (CLA), in 12 cream cheese batches with different prebiotic (inulin) and probiotic (Bifidobacterium animalis and Lactobacillus acidophilus) concentrations stored for 45 days at 8 ± 0.5 °C and analyzed every 15 days. A Central Rotational Composite Design was used to establish the prebiotic and probiotic concentrations. The effect of the pre- and probiotic concentrations on CLA contents (mg g⁻¹ of fat) was verified by plotting the response surface. The linear, quadratic, and interaction coefficients of the multiple linear regression for both prebiotic and probiotics were not significant (p > 0.05). No significant changes were observed neither in the lipid profile nor in the CLA content among the different batches, in which fatty acid composition and CLA concentrations were similar to the milk used in the preparation of the cream cheese and also to the control batch.     

Influence of low‐level supplementation of grazing dairy cows with cereals or sugar beet pulp on the concentrations of CLA isomers in milk

BACKGROUND: A wide range of isomer‐specific health benefits have been attributed to conjugated linoleic acids (CLAs). Little information is currently available on the influence of different feed components on the concentration of CLA isomers in ruminant‐derived products. The aims of this study were to compare the effect of cereal mix or dried sugar beet pulp supplementations fed to grazing dairy cows on milk CLA isomeric distribution and to examine the monthly variation of CLA isomers during the grazing season. RESULTS: The concentrations of the isomers t7c9, t10c12 and t10t12 were higher in milk from cows supplemented with cereals (P ≤ 0.001). The milk of beet pulp‐supplemented cows showed higher levels of the isomers t11c13 (P ≤ 0.05), t9t11 (P ≤ 0.001) and t7t9 (P ≤ 0.01). Total CLA as well as seven other minor isomers were not significantly affected by the concentrate type. Monthly variations occurred extensively for almost all detected isomers. Total CLAs showed the highest values at the beginning of the grazing season and in autumn in coincidence with plant regrowth. CONCLUSION: These results show that even at low levels, supplement types can significantly influence the concentration of some CLA isomers in milk fat; an indication is given of the possibility to manipulate the animals' diet to change the CLA isomeric profile in ruminant‐derived products. Copyright © 2010 Society of Chemical Industry     

Changes in degradation and phosphorylation level of titin in three ovine muscles during postmortem

The objective of this study was to investigate the degradation of titin and its phosphorylation level in three muscles from sheep. The MFI and pH from the longissimus lumborum (LL), semimembranosus (SM) and psoas major (PM) muscles were measured at 30 min, 1, 2, 7, 14, 21 and 28 days postmortem. Myofibrillar proteins were extracted, separated by SDS‐PAGE and quantified by phosphor‐specific staining. Phosphorylation of titin was predicted by Pro‐Q Diamond‐SYPRO Ruby staining. Two days after exsanguination, the pH and MFI of the PM were higher than those of the LL and SM muscles (P < 0.05). The sarcomere length of the PM muscle was also longer than that of the LL and SM muscle (P < 0.05). PM muscles had a highest phosphorylation level (P < 0.05) at 0.5 h postmortem and showed the greatest degree of titin degradation over 28 days. This suggests that phosphorylation of titin might accelerate its degradation.