This paper deals with the theoretical derivation of the modified Haldane model of the substrate inhibition in the microbial growth processes. Based on the biological concepts of substrate-receptor complex working mechanisms, a new microbial kinetics of N-fold multiplex substrate inhibition and its generalization has been considered theoretically, which is natural expansion of the simple substrate inhibition mechanism in the enzyme reaction. As a result, the modified Haldane model of the substrate inhibition turns out to be a well-designed four-parameter kinetic model with a biological constant of the total substrate inhibition concentration.
An experiment was conducted to investigate the effect of microbial phytase (Natuphos$^{\circledR}$) supplementation, individually and in combination with carbohydrase enzyme complex (composed of enzymes targeted to SBM dietary components such as $\alpha$-galactosides and galactomannans; ENDO-POWER$^{\circledR}$) to corn-soy basis diet with low nutrient levels on growth performance and nutrient digestibility of growing pigs. A total of 48 crossbred weaned pigs (Landrace${\times}$Yorkshire${\times}$Duroc), 29.1$\pm$0.14 kg of initial body weight, were randomly allotted to four dietary treatments, based on weight and age, according to a Randomized Complete Block Design. There were three pens per treatment and 4 pigs per pen. The dietary treatments were 1) CON (control diet with 3,380 kcal/kg of metabolizable energy, 18.96% of crude protein, 1.10% of lysine, 0.75% of calcium and 0.35% of available phosphorus), 2) LP+NTPS (CON diet with 0.15% unit lower available P levels+0.1% phytase (500 FTU/kg; Natuphos$^{\circledR}$)), 3) LEL+ENP (CON diet with 3.0% unit lower ME and lysine levels + 0.1% carbohydrase enzyme complex (ENDO-POWER$^{\circledR}$), and 4) LPEL+ENZ (CON diet with 0.15% unit lower available P levels and 3.0% unit lower ME and lysine levels+0.1% ENDO-POWER$^{\circledR}$ and 0.1% Natuphos$^{\circledR}$ (500 FTU/kg). There was no significant difference (p〉0.05) in average daily gain (ADG), average daily feed intake (ADFI) and feed conversion ratio (FCR) among dietary treatments during the whole experimental period (0 to 4 weeks). Apparent digestibility of gross energy was greater in LP+NTPS and LPEL+ENZ groups than in the LEL+ENP (p<0.05). Apparent digestibility of phosphorus was greater in LP+NTPS than in LEL+ENP (p<0.05). Dry matter excretion was lowest in LPEL+ENZ and phosphorus excretion was lowest in LP+NTPS (p<0.05). Overall, pigs fed on LPEL+ENZ group tended to have better nutrient digestibility (dry matter, gross energy, crude protein and phosphorus) than pigs fed on control group. All dietary enzyme treatment groups showed lower feed cost/body weight gain of pigs than control group. In conclusion, the results from the present study suggest that the simultaneous inclusion of phytase and carbohydrase enzyme complex to diets is advantageous with respect to reducing nutrient excretion of growing pigs and may contribute to increased economic return when added to corn-soy based growing pig diets.
Ruminant animals develop a diverse and sophisticated microbial ecosystem for digesting fibrous feedstuffs. Plant cell walls are complex and their structures are not fully understood, but it is generally believed that the chemical properties of some plant cell wall compounds and the cross-linked three-dimensional matrix of polysaccharides, lignin and phenolic compounds limit digestion of cell wall polysaccharides by ruminal microbes. Three adaptive strategies have been identified in the ruminal ecosystem for degrading plant cell walls: production of the full slate of enzymes required to cleave the numerous bonds within cell walls; attachment and colonization of feed particles; and synergetic interactions among ruminal species. Nonetheless, digestion of fibrous feeds remains incomplete, and numerous research attempts have been made to increase this extent of digestion. Exogenous fibrolytic enzymes (EFE) have been used successfully in monogastric animal production for some time. The possibility of adapting EFE as feed additives for ruminants is under intensive study. To date, animal responses to EFE supplements have varied greatly due to differences in enzyme source, application method, and types of diets and livestock. Currently available information suggests delivery of EFE by applying them to feed offers the best chance to increase ruminal digestion. The general tendency of EFE to increase rate, but not extent, of fibre digestion indicates that the products currently on the market for ruminants may not be introducing novel enzyme activities into the rumen. Recent research suggests that cleavage of esterified linkages (e.g., acetylesterase, ferulic acid esterase) within the plant cell wall matrix may be the key to increasing the extent of cell wall digestion in the rumen. Thus, a crucial ingredient in an effective enzyme additive for ruminants may be an as yet undetermined esterase that may not be included, quantified or listed in the majority of available enzyme preparations. Identifying these pivotal enzyme(s) and using biotechnology to enhance their production is necessary for long term improvements in feed digestion using EFE. Pretreating fibrous feeds with alkali in addition to EFE also shows promise for improving the efficacy of enzyme supplements.
Polanski-Cordovano, Grace;Romano, Lea;Marotta, Lauren L.C.;Jacob, Serena;Hoo, Jennifer Soo;Tartaglia, Elena;Asokan, Deepa;Kar, Simkie;Demain, Arnold L.
Journal of Microbiology and Biotechnology
/
v.23
no.5
/
pp.656-660
/
2013
Pseudomonas fluorescens strain CL0145A was discovered at the New York State Museum Field Research Laboratory as an effective agent against the environmentally destructive zebra mussel, which has contaminated US waters. Dried cells of the microbe are being commercialized as an environmentally friendly solution to the problem. We found that antibiotic activity against the Gram-positive bacterium Bacillus subtilis is produced and excreted by this strain. We have carried out studies to optimize production of the antibiotic. Studies were begun in a complex corn meal medium. Activity was found in both cells and culture supernates and was maximal after one day of fermentation. Static fermentation conditions were found to be superior to shaken culture. Production of extracellular antibiotic in complex medium was found to be dependent on the content of sucrose and enzyme-hydrolyzed casein. Indeed, production was greater in sucrose plus enzyme-hydrolyzed casein than in the complex medium. Of a large number of carbon sources studied as improvements over sucrose, the best was glycerol. An examination of nitrogen sources showed that production was improved by replacement of enzyme-hydrolyzed casein with soy hydrolysates. Production in the simple glycerol-Hy-Soy medium was not improved by addition of an inorganic salt mixture or by complex nitrogen sources, with the exception of malt extract. In an attempt to keep the medium more defined, we studied the effect of amino acids and vitamins as replacements for malt extract. Of 21 amino acids and 7 vitamins, we found tryptophan, glutamine, biotin, and riboflavin to be stimulatory. The final medium contained glycerol, Hy-Soy, tryptophan, glutamine, biotin, and riboflavin.
Rumen microbiology research has undergone several evolutionary steps: the isolation and nutritional characterization of readily cultivated microbes; followed by the cloning and sequence analysis of individual genes relevant to key digestive processes; through to the use of small subunit ribosomal RNA (SSU rRNA) sequences for a cultivation-independent examination of microbial diversity. Our knowledge of rumen microbiology has expanded as a result, but the translation of this information into productive alterations of ruminal function has been rather limited. For instance, the cloning and characterization of cellulase genes in Escherichia coli has yielded some valuable information about this complex enzyme system in ruminal bacteria. SSU rRNA analyses have also confirmed that a considerable amount of the microbial diversity in the rumen is not represented in existing culture collections. However, we still have little idea of whether the key, and potentially rate-limiting, gene products and (or) microbial interactions have been identified. Technologies allowing high throughput nucleotide and protein sequence analysis have led to the emergence of two new fields of investigation, genomics and proteomics. Both disciplines can be further subdivided into functional and comparative lines of investigation. The massive accumulation of microbial DNA and protein sequence data, including complete genome sequences, is revolutionizing the way we examine microbial physiology and diversity. We describe here some examples of our use of genomics- and proteomics-based methods, to analyze the cellulase system of Ruminococcus flavefaciens FD-1 and explore the genome of Ruminococcus albus 8. At Illinois, we are using bacterial artificial chromosome (BAC) vectors to create libraries containing large (>75 kbases), contiguous segments of DNA from R. flavefaciens FD-1. Considering that every bacterium is not a candidate for whole genome sequencing, BAC libraries offer an attractive, alternative method to perform physical and functional analyses of a bacterium's genome. Our first plan is to use these BAC clones to determine whether or not cellulases and accessory genes in R. flavefaciens exist in clusters of orthologous genes (COGs). Proteomics is also being used to complement the BAC library/DNA sequencing approach. Proteins differentially expressed in response to carbon source are being identified by 2-D SDS-PAGE, followed by in-gel-digests and peptide mass mapping by MALDI-TOF Mass Spectrometry, as well as peptide sequencing by Edman degradation. At Ohio State, we have used a combination of functional proteomics, mutational analysis and differential display RT-PCR to obtain evidence suggesting that in addition to a cellulosome-like mechanism, R. albus 8 possesses other mechanisms for adhesion to plant surfaces. Genome walking on either side of these differentially expressed transcripts has also resulted in two interesting observations: i) a relatively large number of genes with no matches in the current databases and; ii) the identification of genes with a high level of sequence identity to those identified, until now, in the archaebacteria. Genomics and proteomics will also accelerate our understanding of microbial interactions, and allow a greater degree of in situ analyses in the future. The challenge is to utilize genomics and proteomics to improve our fundamental understanding of microbial physiology, diversity and ecology, and overcome constraints to ruminal function.
Ruminants possess the capacity to digest very large amounts of starch. However, in many cases diets approach 60% starch and even small inefficiencies present opportunities for energetic losses. Ruminal starch digestion is typically 75-80% of starch intake. On average, 35-60% of starch entering the small intestine is degraded. Of the fraction that escapes small-intestinal digestion, 35-50% is degraded in the large intestine. The low digestibility in the large intestine and the inability to reclaim microbial cells imposes a large toll on post-ruminal digestive efficiency. Therefore, digestibility in the small intestine must be optimized. The process of starch assimilation in the ruminant is complex and remains an avenue by which increases in production efficiency can be gained. A more thorough description of these processes is needed before we can accurately predict digestion occurring in the small intestine and formulate diets to optimize site of starch digestion.
This study explores the environmentally innovative and low-impact technology, a zero food waste system (ZFWS) that utilizes food waste and converts it into composts or biofuels and curtails carbon emissions. The ZFWS not just achieves food waste reductions but recycles food waste into fertilizer. Based on a fermentation-extinction technique using bio wood chips, the ZFWS was employed in a field experiment of the system installed in a large-scale apartment complex, and the performance of the system was examined. The on-site ZFWS consisted of three primary parts: 1) a food waste slot into which food waste was injected; 2) a fermentation-extinction reactor where food waste was mixed with bio wood chips made up of complex enzyme and aseptic wood chips; and 3) deodorization equipment in which an ultraviolet and ozone photolysis method was employed. The field experiment showed that food waste injected into the ZFWS was reduced by 94%. Overall microbial activity of the food waste in the fermentation-extinction reactor was measured using adenosine tri-phosphate (ATP), and the degradation rate of organic compounds, referred to as volatile solids, increased with ATP concentration. The by-products generated from ZFWS comply with the national standard for organic fertilizer.
An experiment was conducted to investigate the effects of microbial phytase ($Natuphos^{(R)}$) supplementation in combination with carbohydrases (composed of enzymes targeted to soybean meal (SBM) dietary components such as $\alpha$-galactosides and galactomannans; $Endo-Power^{(R)}$) to corn-soybean meal based diet (CSD) and complex diet (CD) with a partial replacement of SBM with rape seed meal (RSM) and cotton seed meal (CSM) on growth performance and nutrient digestibility of growing pigs. A total of 168 growing pigs averaging $13.18{\pm}1.77kg$ of initial body weight was arranged as a $2{\times}2$ factorial design with main effects of diet types (corn-SBM based diet (CSD) and complex diets (CD; 5% of SBM was replaced with 2.5% of RSM and 2.5% of CSM in diet for phase I (0 to 3 weeks) and 6% of SBM was replaced with 3% of RSM and 3% of CSM in diet for phase II (4 to 7 weeks))) and enzyme supplementation (none and 0.1% of phytase (500 FTU/kg diet) and 0.1% of carbohydrases). The diet with enzyme application were formulated to have a 0.18% unit lower aP than diets without enzyme application. Each treatment had three replicates with 14 pigs per replicate. To determine supplementation effect of phytase and carbohydrases on ileal amino acid digestibility of SBM, RSM and CSM, a total of 18 T-cannulated pigs (initial body weight; $13.52{\pm}1.24kg$) were assigned to six dietary treatments in the present study. Dietary treatments in metabolic trial included 1) SBM diet, 2) SBM diet+with enzymes (phytase (500 FTU/kg) and carbohydrases at 0.1%, respectively), 3) CSM diet, 4) CSM diet+enzymes, 5) RSM diet and 6) RSM diet+enzymes. During whole experimental period (0 to 7 wks), there was no difference in growth performance between diets (CSD and CD). However, dietary phytase and carbohydrases supplementation significantly improved gain/feed ratio (G:F) of growing pigs. During the phase II (4-7 weeks), dietary phytase and carbohydrases supplementation significantly improved all fecal nutrient digestibilities (Dry matter (DM), gross energy (GE), crude protein (CP), crude fat (CF), calcium (Ca) and phosphorus (P)). Dietary phytase and carbohydrases supplementation improved significantly overall ileal amino acid digestibilities of SBM, RSM and CSM based diets (p<0.05). The simultaneous inclusion of phytase and carbohydrases in both of CSD and CD reduced feed cost per kg body weight gain (FCG). Also, results suggest that 2.5 to 3% of RSM and CSM, respectively, might be used as a protein source in growing pig diets without having an adverse effect on the growth performance and nutrient digestibility and simultaneous phytase and carbohydrases addition improves nutritional value of SBM, RSM and CSM by improving ileal amino acid digestibilities.
The human gastrointestinal (GI) tract houses a diverse microbial community, known as the gut microbiome comprising bacteria, viruses, fungi, and protozoa. The gut microbiome plays a crucial role in maintaining the body's equilibrium and has recently been discovered to influence the functioning of the central nervous system (CNS). The communication between the nervous system and the GI tract occurs through a two-way network called the gut-brain axis. The nervous system and the GI tract can modulate each other through activated neuronal cells, the immune system, and metabolites produced by the gut microbiome. Extensive research both in preclinical and clinical realms, has highlighted the complex relationship between the gut and diseases associated with the CNS, such as Alzheimer's disease, Parkinson's disease, and amyotrophic lateral sclerosis. This review aims to delineate receptor and target enzymes linked with gut microbiota metabolites and explore their specific roles within the brain, particularly their impact on CNS-related diseases.
Edible films containing antimicrobial agents can be used as safe alternatives to preserve food products. Essential oils are well-recognized antimicrobials. However, their low water solubility, volatility and high sensitivity to oxygen and light limit their application in food preservation. These limitations could be overcome by embedding these essential oils in complexed product matrices exploiting the encapsulation efficiency of β-cyclodextrin. This study focused on the maximization of β-cyclodextrin production using cyclodextrin glucanotransferase (CGTase) and the evaluation of its encapsulation efficacy to fabricate edible antimicrobial films. Response surface methodology (RSM) was used to optimize CGTase production by Brevibacillus brevis AMI-2 isolated from mangrove sediments. This enzyme was partially purified using a starch adsorption method and entrapped in calcium alginate. Cyclodextrin produced by the immobilized enzyme was then confirmed using high performance thin layer chromatography, and its encapsulation efficiency was investigated. The clove oil/β-cyclodextrin inclusion complexes were prepared using the coprecipitation method, and incorporated into chitosan films, and subjected to antimicrobial testing. Results revealed that β-cyclodextrin was produced as a major product of the enzymatic reaction. In addition, the incorporation of clove oil/β-cyclodextrin inclusion complexes significantly increased the antimicrobial activity of chitosan films against Staphylococcus aureus, Staphylococcus epidermidis, Salmonella Typhimurium, Escherichia coli, and Candida albicans. In conclusion, B. brevis AMI-2 is a promising source for CGTase to synthesize β-cyclodextrin with considerable encapsulation efficiency. Further, the obtained results suggest that chitosan films containing clove oils encapsulated in β-cyclodextrin could serve as edible antimicrobial food-packaging materials to combat microbial contamination.
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