Comparative Genomic Analysis of Limosilactobacillus Pontis Reveals Genetic Characteristics of High GC Content and Potential Niches Adaptations

Limosilactobacillus pontis is a species of lactic acid bacteria (LAB) found in fermented milk, sourdough and broiler chickens gastrointestinal tract and so on. However, the evolutionary strategies and genomic characteristics of the species remain unknown, which limits its application. In this study, whole genome sequencing was combined with a comparative genomic approach to investigate genomic characteristics and evolutionary strategies of L. pontis; this includes three published genomic sequences and two strains isolated from fermented milk in Inner Mongolia, China. The mean genome size and GC content of L. pontis was 1.70 Mb and 53.06%, respectively. Within the LAB L. pontis has a high GC content. The phylogenetic tree based on 1,281 core genomes showed that strains from the same sources aggregated together in clusters. Genome information, average nucleotide identity values, and phylogenetic relationships amongst L. pontis from different sources indicated that strains have potential niche adaptability. Functional genomic aspects, GT2 and GT4 (glycosyltransferases, GTs) involved in the synthesis of cellulose and sucrose were the family with the largest number of carbohydrate enzymes in L. pontis, particularly strains isolated from fermented milk. It is worth mentioning that the ability of L. pontis to produce bacteriocin may increase its application potential. This study provides new insight into the genetic characteristics and potential niche adaptations of L. pontis.

314 Carbohydrases Enzymes Improve the Productive Performance of Grazing Nellore Bulls Fed Different Nutritional Plans During the Rainy Season

The aim with this trial was to evaluate two nutritional plans with the addition of dosages of carbohydrases enzyme blend on productive performance of grazing Nellore bulls in rainy season. One hundred and two contemporary Nellore steers (BW = 294.10 ± 3.35 kg) were equally distributed in six paddocks (17 steers/paddock) following a completely randomized design in a 2x3 factorial arrangement, being the factors two nutritional plans: mineral-proteic supplementation (Fosbovi® Proteico 30; PS) offered at 0,1% BW or mineral-protein-energetic supplementation (Fosbovi® Proteico-Energético 25; PES) offered at 0.3% BW); and three levels of enzymatic carbohydrases blend (0, 4,75 or 9,50 g/animal/day). The enzymatic blend was mainly composed by beta-glucanase and xylanase enzymes (Ronozyme® VP and Ronozyme® WX); Both enzymes and supplemented were provided by DSM Nutritional Products Brasil S.A. Supplement intake and animal weighting were carried out daily and individually by an automatic scale system installed in each paddock. Data were analyzed using PROC MIXED of SAS and means were compared by polynomial regression. Animals that received PES had greater ADG (0.896 vs. 0.775 kg/day; P < 0.001) compared to those fed PS. Intermediary level (4.75 g/day) of carbohydrate enzymes also presented the highest ADG in PES compared to 0.0 level (0.941 vs. 0.896 kg/day; P = 0.006). Still, higher enzyme levels (9.5 g/day) presented greater ADG on PS, when compared to 0.0 level (0.852 vs. 0.775 kg/day, P = 0.006). PES animals presented greater intake, compared to PS (1.112 vs. 0.432 kg/day, P < 0.001). According to polynomial regression, it is recommended to include 4.75 g/day of these carbohydrases enzymes in PS and 4.39 g/day in PES for growing Nellore bulls in rainy season.

Diversity of Weissella confusa in Pozol and Its Carbohydrate Metabolism

The genus Weissella is composed of a group of Gram-positive facultative anaerobe bacteria with fermentative metabolism. Strains of this genus have been isolated from various ecological niches, including a wide variety of fermented cereal foods. The present study aimed to determine the relative abundance and fermentation capabilities of Weissella species isolated from pozol, a traditional maya product made of lime-cooked (nixtamalized) fermented maize. We sequenced the V3-V4 regions of 16S rDNA; Weissella was detected early in the fermentation process and reached its highest relative abundance (3.89%) after 3 h of culture. In addition, we evaluated five Weissella strains previously isolated from pozol but reported as non-amylolytic, to define alternative carbon sources such as xylan, xylooligosaccharides, and sucrose. While no growth was observed on birch xylan, growth did occur on xylooligosaccharides and sucrose. Strains WcL17 and WCP-3A were selected for genomic sequencing, as the former shows efficient growth on xylooligosaccharides and the latter displays high glycosyltransferase (GTF) activity. Genomes of both strains were assembled and recorded, with a total of 2.3 Mb in 30 contigs for WcL17 and 2.2 Mb in 45 contigs for WCP-3a. Both strains were taxonomically assigned to Weissella confusa and genomic analyses were performed to evaluate the gene products encoding active carbohydrate enzymes (CAZy). Both strains have the gene content needed to metabolize sucrose, hemicellulose, cellulose, and starch residues, all available in pozol. Our results suggest that the range of secondary enzymatic activity in Weissella confusa strains confer them with wide capabilities to participate in fermentative processes of natural products with heterogeneous carbon sources.

Knockdown of β-N-acetylglucosaminidase 2 Impairs Molting and Wing Development in Lasioderma serricorne (Fabricius)

β-N-acetylglucosaminidases (NAGs) are carbohydrate enzymes that degrade chitin oligosaccharides into N-acetylglucosamine monomers. This process is important for chitin degradation during insect development and metamorphosis. We identified and evaluated a β-N-acetylglucosaminidase 2 gene (LsNAG2) from the cigarette beetle, Lasioderma serricorne (Fabricius). The full-length open reading frame of LsNAG2 was 1776 bp and encoded a 591 amino acid protein. The glycoside hydrolase family 20 (GH20) catalytic domain and an additional GH20b domain of the LsNAG2 protein were highly conserved. Phylogenetic analysis revealed that LsNAG2 clustered with the group II NAGs. Quantitative real-time PCR analyses showed that LsNAG2 was expressed in all developmental stages and was most highly expressed in the late larval and late pupal stages. In the larval stage, LsNAG2 was predominantly expressed in the integument. Knockdown of LsNAG2 in fifth instar larvae disrupted larval–pupal molting and reduced the expression of four chitin synthesis genes (trehalase 1 (LsTRE1), UDP-N-acetylglucosamine pyrophosphorylase 1 and 2 (LsUAP1 and LsUAP2), and chitin synthase 1 (LsCHS1)). In late pupae, LsNAG2 depletion resulted in abnormal adult eclosion and wing deformities. The expression of five wing development-related genes (teashirt (LsTSH), vestigial (LsVG), wingless (LsWG), ventral veins lacking (LsVVL), and distal-less (LsDLL)) significantly declined in the LsNAG2-depleted beetles. These findings suggest that LsNAG2 is important for successful molting and wing development of L. serricorne.

Cis-nonPro Peptides: Genuine Occurrences and their Functional Roles

Cis-nonPro peptides, a very rare feature in protein structures, are of considerable importance for two opposite reasons. On one hand, their genuine occurrences are mostly found at sites critical to biological function, from the active sites of carbohydrate enzymes to rare adjacent-residue disulfide bonds. On the other hand, a cis-nonPro can easily be misfit into weak or ambiguous electron density, which has led to a high incidence of unjustified cis-nonPro over the last decade. This paper uses the greatly expanded crystallographic data and newly stringent quality-filtering to identify the genuine occurrences and survey both individual examples and broad patterns of their functionality. The accompanying paper describes the problem of cis-nonPro over-use, including its causes, validation, and correction.We explain the procedure developed to identify genuine cis-nonPro examples with almost no false positives, including the new observation that peptides with a glycine on one side or the other need extra care to avoid mis-assignment as cis-nonPro. We then survey a sample of the varied functional roles and structural contexts of cis-nonPro, emphasizing aspects not previously covered systematically: the preferred occurrence at β-strand ends in TIM barrel structures, the concentration of occurrence in proteins that process, bind, or contain carbohydrates, and the resulting complications in defining a simple occurrence frequency.

Evaluation of anti-hyperglycemic and anti-hyperlipidemic effects of Naravelia zeylanica in streptozotocin- induced diabetic rats

<p><em>Naravelia zeylanica</em> DC (Ranunculaceae), a woody climber, have been used from ancient times to treat various ailments like rheumatoid arthritis, skin diseases, wound and ulcer. The present study was designed to investigate the anti-hyperglycemic activity of methanolic extract of <em>Naravelia zeylanica</em> (NZYM) using experimental diabetic model. Diabetes was induced in<strong> </strong>wistar rats by a single dose of streptozotocin (55 mg/kg i.p.) (STZ) and treated with NZYM at doses of 100 and 200 mg/kg for 45 days. Glibenclamide (5 mg/kg b.w.) was used as standard drug. Blood glucose and body weight were monitored at regular intervals and the levels of serum insulin, lipid and the carbohydrate metabolic enzyme in the liver were measured at the end of the study. Oral administration of NZYM and glibenclamide significantly reduced the blood glucose level (<em>p&lt;0.05</em>), with increased serum insulin and significant alteration in lipid profiles and liver carbohydrate enzymes (<em>p&lt;0.05</em>) after 45 days. Furthermore, the biochemical parameters correlated with the histopathological changes in the pancreas of STZ-induced diabetic rats, which structurally proved the efficacy of NZYM. The findings suggest that NZYM possess anti-hyperglycemic activity and anti-hyperlipidemic properties and restored STZ-induced pancreatic damage in diabetic rats. NZYM might therefore have a beneficial effect in treatment of diabetes mediated through regulation of carbohydrate metabolic enzyme activities. <strong></strong></p>