Hui Gyu Park

Synthesis of conjugated linoleic acid by human-derived Bifidobacterium breve LMC 017: utilization as a functional starter culture for milk fermentation.

This study was performed to discover bifidobacteria isolated from human intestines that optimally convert linoleic acid (LA) to conjugated linoleic acid (CLA) and to optimize the culture conditions of milk fermentation. One hundred and fifty neonatal bifidobacteria were screened for CLA-producing ability, and Bifidobacterium breve LMC 017 was selected as it showed about 90% conversion of free LA in MRS broth. The selected strain showed resistance at 0.5% LA in microaerophillic conditions. When monolinolein (LA 90%) was used as a substrate for CLA production, the conversion rate was lower compared to free LA, but the growth rate was unaffected during the milk fermentation. There was no significant difference in CLA production between aerobic and anaerobic conditions, and little decline in CLA was shown after the maximal CLA level had been reached. CLA production increased by 80% with 24 h of incubation in milk containing additional skim milk (5%), where the proteins may have facilitated the production of CLA by enhancing the interaction of substrate with the bacteria. CLA production did not decline after 9 h of fermentation and an additional 12 weeks of storage with other commercial starters. This demonstrates the possibility of using this strain as a costarter in the production of CLA-enriched yogurt.

Characterization of conjugated linoleic acid production by Bifidobacterium breve LMC 520.

This study was performed to characterize the CLA production ability of a bacterial strain, Bifidobacterium breve LMC 520, which can actively convert linoleic acid (LA) to cis-9,trans-11 conjugated linoleic acid (CLA), a major isomer derived from microbial enzymatic conversion. The culture conditions were optimized to improve CLA production under the aerobic conditions. B. breve LMC 520 was tested with different amounts of LA in varied culture conditions, such as air, additives, and pH. A maximal level of CLA production (up to 90% of substrate) was obtained after 24 h of incubation in culture medium containing 1 mM LA at pH 5.5 and under anaerobic conditions. There was no decline in the CLA level with prolonged incubation until 48 h. When the effect of pre-incubation with LA on CLA production was tested, there was no significant difference between the CLA-producing activity of pre-incubated and untreated bacteria at the third passage but there was a significant reduction in CLA production by the pre-incubated cells after the fourth passage. These results demonstrate that the CLA-producing activity of B. breve LMC 520 could be maximized by numerous environmental factors. The data also indicate its potential for increasing CLA accumulation in dairy products when B. breve LMC 520 is used as a functional starter culture.

Dietary conjugated linoleic acid increases endurance capacity of mice during treadmill exercise.

This study was designed to evaluate the effect of dietary conjugated linoleic acid (CLA) on endurance exercise and availability of metabolic substrates in mice. Four-week-old male BALB/c mice were randomized to a control group (normal diet) and a CLA group (normal diet + 1% CLA). Each animal group received its respective diet for 10 weeks and was subjected to forced running on a treadmill system to estimate their maximum endurance capacity at the end of the experiment. All mice were immediately sacrificed after prolonged exercise, and the major metabolic substrates were measured in serum and liver. The body weights of mice in the CLA group were lower than those of the control group after the 10 weeks. The maximum running time in CLA-fed mice was significantly longer, by 26%, compared to that of the control mice. Dietary CLA decreased the serum concentrations of triglycerides, nonesterified fatty acids, and urea nitrogen and significantly reduced the consumption of liver glycogen. The present study demonstrated that dietary CLA enhances the endurance capacity of mice by increasing fat utilization and reducing the consumption of stored liver glycogen as substrates for energy metabolism.

Utilization of Monolinolein as a Substrate for Conjugated Linoleic Acid Production by Bifidobacterium breve LMC 520 of Human Neonatal Origin

This study was designed to isolate bifidobacteria from human intestines that efficiently converts monolinolein, a monoglyceride form of linoleic acid, into conjugated linoleic acid (CLA), as well as to optimize culture conditions for improving CLA production during milk fermentation. Among 150 screened neonatal bifidobacteria, Bifidobacterium breve LMC 520 showed the highest CLA-producing ability and was tested with different types of fat substrates at various concentrations to determine the optimal conditions for CLA production. Monolinolein was tested as a substrate for CLA production. The incubation time optimized for CLA production was 24 h, and CLA production was proportionally increased with monolinolein concentration. The incubation of LMC 520 with commercial starter strains caused minimal reduction in CLA production. Our results demonstrate that the CLA-producing ability of B. breve LMC 520 could offer beneficial effects when utilized as a starter culture for the development of functional dairy products.

Production of a Conjugated Fatty Acid by Bifidobacterium breve LMC520 from α-Linolenic Acid: Conjugated Linolenic Acid (CLnA)

This study was performed to characterize natural CLnA isomer production by Bifidobacterium breve LMC520 of human origin in comparison to conjugated linoleic acid (CLA) production. B. breve LMC520 was found to be highly active in terms of CLnA production, of which the major portion was identified as cis-9,trans-11,cis-15 CLnA isomer by GC-MS and NMR analysis. B. breve LMC520 was incubated for 48 h using MRS medium (containing 0.05% L-cysteine · HCl) under different environmental conditions such as atmosphere, pH, and substrate concentration. The high conversion rate of α-linolenic acid (α-LNA) to CLnA (99%) was retained up to 2 mM α-LNA, and the production was proportionally increased nearly 7-fold with 8 mM by the 6 h of incubation under anaerobic conditions at a wide range of pH values (between 5 and 9). When α-LNA was compared with linoleic acid (LA) as a substrate for isomerization by B. breve LMC520, the conversion of α-LNA was higher than that of LA. These results demonstrated that specific CLnA isomer could be produced through active bacterial conversion at an optimized condition. Because many conjugated octadecatrienoic acids in nature are shown to play many positive roles, the noble isomer found in this study has potential as a functional source.

Functional comparison of esterified and free forms of conjugated linoleic acid in high-fat-diet-induced obese C57BL/6J mice.

This study investigated the effects of dietary conjugated linoleic acid (CLA), in the form of free fatty acid (FFA-CLA) or triacylglycerol (TG-CLA), on serum and liver lipid composition and gene expression associated with lipogenesis and β-oxidation in high-fat-diet (HFD)-induced obese C57BL/6J mice. Animals were fed a control diet, HFD, HFD supplemented with 2% FFA-CLA, or HFD supplemented with 2% TG-CLA for 8 weeks. Supplementation with both forms of CLA significantly reduced the weights of whole body and adipose tissue and was positively associated with significant liver enlargement. Both forms of CLA significantly decreased serum TG concentration, but had no effect on total cholesterol levels, which were increased in mice fed HFD. There was a prominent increase in serum alanine aminotransferase (ALT) levels in mice that received either form of CLA. TG accumulation and lipogenic gene expression, including the expression of genes for fatty acid synthase (FAS), acetyl-coenzyme A carboxylase (ACC), and malic enzyme, were significantly lower in the livers of mice that received TG-CLA as compared to FFA-CLA. The gene expressions of sterol regulatory element binding protein-1c (SREBP-1c) in both liver and adipose tissue were suppressed in mice that were fed either form of CLA as compared to the mice fed HFD alone, whereas there were no increases in the levels of expression of β-oxidation-related genes. These findings demonstrated that free and esterified forms of CLA have differing effects on liver and adipose tissue lipogenesis.

Production of Conjugated Linoleic Acid (CLA) by Bifidobacterium breve LMC520 and Its Compatibility with CLA-Producing Rumen Bacteria

This study was performed to characterize the ability of an active Bifidobacterium strain to produce conjugated linoleic acid (CLA) and to test its possible utilization as a probiotic compatible to the ruminal condition. Bifidobacterium breve LMC520 can actively convert linoleic acid (LA) to cis-9,trans-11-CLA, which is a major isomer derived from microbial conversion. LMC520 showed reasonable tolerance under acidic conditions (pH 2.5 with 1% pepsin) and in the presence of oxgall (0-3%). The growth and CLA production of LMC520 were tested under ruminal conditions and compared with those of Butyrivibrio fibrisolvens A38, which is a major CLA producer in the rumen as an intermediate in the biohydrogenation (BH) process. LMC520 converted 15% of LA to CLA under ruminal conditions, which was 2 times higher activity than that of A38, and there was no decline in CLA level during prolonged incubation of 48 h. The BH activity of LMC520 was comparable to that of A38. When LMC520 was cocultured with A38, even with slight decrease of CLA due to high BH activity by A38, but the level of CLA was maintained by the high CLA-producing activity of LMC520. This comparative study shows the potential of this strain to be applied as a functional probiotic not only for humans but also for ruminants as well as to increase CLA production.

Characterizations of environmental factors in conjugated linoleic acid production by mixed rumen bacteria.

Conjugated linoleic acid (CLA) production by rumen bacteria is closely related to biohydrogenation of linoleic acid (LA) and affected by various environmental factors. Ruminal biohydrogenation and isomerization were characterized in view of incubation conditions using a mixed culture of microorganisms obtained from surgically prepared cows. Free-floating bacteria (FFB) produced more CLA than particle-attached bacteria (PAB). Some major factors affecting the ruminal environment such as diet, concentrations of fat substrates, incubation time, pre-incubation, and the presence of glucose in the medium were found to be important determinants for the ruminal production of CLA and in a close relationship with biohydrogenation. The mixed bacterial culture, which was pre-exposed to LA, produced more CLA than an unexposed control in a medium containing 30% rumen fluid. The rate of conversion of fat substrate (LA) to hydrogenated products (trans-C18:1, C18:0) was negatively correlated with the initial LA concentration. Overall, the present study showed that CLA accumulation can be increased by modification of diet-induced environmental conditions, which affect changes in ruminal microflora.

Effect of High Hydrostatic Pressure on the Production of Conjugated Fatty Acids and Trans Fatty Acids by Bifidobacterium breve LMC520

This study was performed to investigate the effect of high hydrostatic pressure (HHP) on the conversion of linoleic acid, conjugated linoleic acid (CLA), and α-linolenic acid (α-LNA) as substrates by Bifidobacterium breve LMC520 and to optimize the HHP condition. Cell mixture were tested under HHP in a variety of conditions such as temperature, time, pressure, and pre- or post-treatment with substrates. The cis-9,trans-11 CLA producing activity of B. breve LMC520 was increased by HHP, whereas trans-9,trans-11 CLA producing activity was decreased. Optimal HHP conditions for the highest CLA production were obtained at 100 MPa for 12 h at 37 °C. Post-treatment groups showed higher conversion activity of substrates than pretreatment groups. Post-treatment groups decreased trans-9,trans-11 CLA and other CLnA, whereas the pretreatment groups increased them. It is concluded that HHP treatment could be an important factor to enhance CLA and CLnA production and for reducing trans-fatty acids.