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Characterization of Feruloyl Esterases Produced by the Four Lactobacillus Species: L. amylovorus, L. acidophilus, L. farciminis and L. fermentum, Isolated from Ensiled Corn Stover.

ABSTRACT: Lactic acid bacteria (LAB) play important roles in silage fermentation, which depends on the production of sufficient organic acids to inhibit the growth of undesirable microorganisms. However, LAB are not able to degrade cellulose and hemicellulose. Bacteria and fibrolytic enzymes are usually used as inoculants to improve the silage quality and digestibility. In the present study, we isolated four Lactobacillus strains (L. amylovorus CGMCC 11056, L. acidophilus CCTCC AB2010208, L. farciminis CCTCC AB2016237 and L. fermentum CCTCC AB2010204) with feruloyl esterase (FAE) activities from ensiled corn stover (CS) by a plate screening assay. The genes encoding FAEs were cloned and hetero-expressed in Escherichia coli. The optimal temperature and pH of these purified enzymes ranged from 45 to 50°C and from 7.0 to 8.0, respectively. They could hydrolyze hydroxycinnamoyl esters in a substrate-specific manner when methyl ferulate, methyl caffeate, methyl ?-coumarate and methyl sinapinate were used as substrates. Moreover, these four FAEs were able to hydrolyze CS to release hydroxycinnamic acids. Furthermore, these strains could degrade hydroxycinnamic esters, and L. amylovorus CGMCC 11056 was the most efficient strain among these four isolates. These results provided a new target for the development of inoculants to improve silage quality and digestibility.

SUBMITTER: Xu Z

PROVIDER: S-EPMC5454770 | biostudies-literature | 2017

REPOSITORIES: biostudies-literature

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Characterization of Feruloyl Esterases Produced by the Four Lactobacillus Species: L. amylovorus, L. acidophilus, L. farciminis and L. fermentum, Isolated from Ensiled Corn Stover.

Xu Zhenshang Z He Huiying H Zhang Susu S Guo Tingting T Kong Jian J

Frontiers in microbiology 20170602

Lactic acid bacteria (LAB) play important roles in silage fermentation, which depends on the production of sufficient organic acids to inhibit the growth of undesirable microorganisms. However, LAB are not able to degrade cellulose and hemicellulose. Bacteria and fibrolytic enzymes are usually used as inoculants to improve the silage quality and digestibility. In the present study, we isolated four Lactobacillus strains (L. amylovorus CGMCC 11056, L. acidophilus CCTCC AB2010 ...[more]

PMID: 28626449

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Project description:IntroductionThe aim of this study was to investigate the effects of adding whole-plant ensiled corn stalks (WECS) to the diet of Holdorbagy geese on their growth performance, serum parameters, and cecal microbiota. Geese farming is an important agricultural practice, and optimizing their diet can contribute to better growth and health outcomes. However, there is limited research on the utilization of WECS as a feed source for geese. Understanding the potential effects of WECS on growth, blood parameters, and cecal microbiota can provide valuable insights into its feasibility and impact on geese farming practices.MethodsA total of 144 six-week-old Holdorbagy geese were randomly assigned to one of three groups: a control group (0% WECS), a group fed 15% WECS and 85% concentrated feed (15% WECS), and a group fed 30% WECS and 70% concentrated feed (30% WECS). The trial period lasted for three weeks, during which the growth performance, serum parameters, and cecal microbiota were assessed.ResultsThe results revealed significant findings in different aspects. Firstly, the feed-to-gain ratio (F/G ratio) of the 15% WECS group was significantly higher than that of the control group (p<0.05), indicating potential challenges in feed efficiency. Additionally, the average daily feed intake (ADFI) of both the 15% and 30% WECS groups was significantly higher than that of the control group (p<0.05), suggesting increased appetite or palatability of the diet containing WECS. In terms of serum parameters, the level of lactate dehydrogenase (LDH) in the 30% WECS group was significantly lower than that in the control group (p<0.05). Moreover, there was a tendency for increasing Fe levels and decreasing Zn levels with higher levels of WECS supplementation, although the differences were not statistically significant (p<0.05). Furthermore, the principal coordinate analysis showed significant differences in the composition of cecal microbiota among the three groups (p < 0.01). The observed_species, Shannon, and Pielou_e indices of the 30% WECS group were significantly higher than those of the 0% and 15% WECS groups (p<0.05), while the Simpson index of the 15% WECS group was significantly lower than that of the control group (p<0.05).DiscussionThe results indicate that the addition of WECS to the geese diet has both positive and negative effects. The study suggests that WECS can be a long-term stable feed source for geese, which can contribute to reducing feeding costs. However, it is important to monitor the amount of WECS added as it can affect the absorption of Zn by geese. Supplementation of Zn in the diet might be necessary to meet the needs of geese. Notably, adding 30% WECS to the diet can increase the richness, evenness, and diversity of the cecal microbiota, indicating potential benefits to gut health. In conclusion, this study highlights the potential of WECS as a feed source for geese. It provides valuable insights into the effects of WECS on growth performance, serum parameters, and cecal microbiota. These findings contribute to optimizing geese farming practices, improving feed utilization, and enhancing overall productivity and well-being of geese. Further research is needed to determine the optimal inclusion level of WECS and to explore strategies for mitigating any negative effects.

Project description:UnlabelledA total of 33 Lactobacillus strains were screened for feruloyl esterase (FE) activity using agar plates containing ethyl ferulate as the sole carbon source, and Lactobacillus fermentum NRRL B-1932 demonstrated the strongest FE activity among a dozen species showing a clearing zone on the opaque plate containing ethyl ferulate. FE activities were monitored using high-performance liquid chromatography with an acetonitrile-trifluoroacetic acid gradient. To produce sufficient purified FE from L. fermentum strain NRRL B-1932 (LfFE), the cDNA encoding LfFE (Lffae) was amplified and cloned by using available closely related genome sequences and overexpressed in Escherichia coli A 29.6-kDa LfFE protein was detected from the protein extract of E. coli BL21(pLysS) carrying pET28bLffae upon IPTG (isopropyl-β-d-thiogalactopyranoside) induction. The recombinant LfFE containing a polyhistidine tag was purified by nickel-nitrilotriacetic acid affinity resin. The purified LfFE showed strong activities against several artificial substrates, including p-nitrophenyl acetate and 4-methylumbelliferyl p-trimethylammoniocinnamate chloride. The optimum pH and temperature of the recombinant LfFE were around 6.5 and 37°C, respectively, as determined using either crude or purified recombinant LfFE. This study will be essential for the production of the LfFE in E. coli on a larger scale that could not be readily achieved by L. fermentum fermentation.ImportanceThe production of feruloyl esterase (FE) from Lactobacillus fermentum NRRL B-1932 reported in this study will have immense potential commercial applications not only in biofuel production but also in pharmaceutical, polymer, oleo chemical, cosmetic additive, and detergent industries, as well as human health-related applications, including food flavoring, functional foods, probiotic agents, preventive medicine, and animal feed. Given the essential role FE plays in the production of hydroxycinnamic acids and ferulic acid, plus the generally regarded as safe status of lactobacilli, which therefore have less regulatory concerns, LfFE from the probiotic L. fermentum reported in this work can be directly used for increased production of high-value hydroxycinnamates and ferulic acid from natural or synthetic carbon sources.

Dataset Information

Characterization of Feruloyl Esterases Produced by the Four Lactobacillus Species: L. amylovorus, L. acidophilus, L. farciminis and L. fermentum, Isolated from Ensiled Corn Stover.

Publications

Characterization of Feruloyl Esterases Produced by the Four <i>Lactobacillus</i> Species: <i>L. amylovorus</i>, <i>L. acidophilus</i>, <i>L. farciminis</i> and <i>L. fermentum</i>, Isolated from Ensiled Corn Stover.

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