Fundamental features of microbial cellulose utilization are examined at successively higher levels of aggregation encompassing the structure and composition of cellulosic biomass, taxonomic diversity, cellulase enzyme systems, molecular biology of cellulase enzymes, physiology of cellulolytic microorganisms, ecological aspects of cellulase-degrading communities, and rate-limiting factors in nature. The methodological basis for studying microbial cellulose utilization is considered relative to quantification of cells and enzymes in the presence of solid substrates as well as apparatus and analysis for cellulose-grown continuous cultures. Quantitative description of cellulose hydrolysis is addressed with respect to adsorption of cellulase enzymes, rates of enzymatic hydrolysis, bioenergetics of microbial cellulose utilization, kinetics of microbial cellulose utilization, and contrasting features compared to soluble substrate kinetics. A biological perspective on processing cellulosic biomass is presented, including features of pretreated substrates and alternative process configurations. Organism development is considered for "consolidated bioprocessing" (CBP), in which the production of cellulolytic enzymes, hydrolysis of biomass, and fermentation of resulting sugars to desired products occur in one step. Two organism development strategies for CBP are examined: (i) improve product yield and tolerance in microorganisms able to utilize cellulose, or (ii) express a heterologous system for cellulose hydrolysis and utilization in microorganisms that exhibit high product yield and tolerance. A concluding discussion identifies unresolved issues pertaining to microbial cellulose utilization, suggests approaches by which such issues might be resolved, and contrasts a microbially oriented cellulose hydrolysis paradigm to the more conventional enzymatically oriented paradigm in both fundamental and applied contexts.

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Microbial cellulose utilization: fundamentals and biotechnology.

Microbial pectinolytic enzymes: A review

Astaxanthin is a carotenoid widely used in salmonid and crustacean aquaculture to provide the pink color characteristic of that species. This application has been well documented for over two decades and is currently the major market driver for the pigment. Additionally, astaxanthin also plays a key role as an intermediary in reproductive processes. Synthetic astaxanthin dominates the world market but recent interest in natural sources of the pigment has increased substantially. Common sources of natural astaxanthin are the green algae Haematococcus pluvialis, the red yeast, Phaffia rhodozyma, as well as crustacean byproducts. Astaxanthin possesses an unusual antioxidant activity which has caused a surge in the nutraceutical market for the encapsulated product. Also, health benefits such as cardiovascular disease prevention, immune system boosting, bioactivity against Helycobacter pylori, and cataract prevention, have been associated with astaxanthin consumption. Research on the health benefits of astaxanthin is very recent and has mostly been performed in vitro or at the pre-clinical level with humans. This paper reviews the current available evidence regarding astaxanthin chemistry and its potential beneficial effects in humans.

Astaxanthin: a review of its chemistry and applications.

Genome-wide transcript profiling was used to monitor signal transduction during yeast pheromone response. Genetic manipulations allowed analysis of changes in gene expression underlying pheromone signaling, cell cycle control, and polarized morphogenesis. A two-dimensional hierarchical clustered matrix, covering 383 of the most highly regulated genes, was constructed from 46 diverse experimental conditions. Diagnostic subsets of coexpressed genes reflected signaling activity, cross talk, and overlap of multiple mitogen-activated protein kinase (MAPK) pathways. Analysis of the profiles specified by two different MAPKs-Fus3p and Kss1p-revealed functional overlap of the filamentous growth and mating responses. Global transcript analysis reflects biological responses associated with the activation and perturbation of signal transduction pathways.

https://science.sciencemag.org/content/sci/287/5454/873.full.pdf

Signaling and Circuitry of Multiple MAPK Pathways Revealed by a Matrix of Global Gene Expression Profiles

The first stage of chocolate production consists of a natural, seven-day microbial fermentation of the pectinaceous pulp surrounding beans of the tree Theobroma cacao. There is a microbial succession of a wide range of yeasts, lactic-acid, and acetic-acid bacteria during which high temperatures of up to 50°C and microbial products, such as ethanol, lactic acid, and acetic acid, kill the beans and cause production of flavor precursors. Over-fermentation leads to a rise in bacilli and filamentous fungi that can cause off-flavors. The physiological roles of the predominant micro-organisms are now reasonably well understood and the crucial importance of a well-ordered microbial succession in cocoa aroma has been established. It has been possible to use a synthetic microbial cocktail inoculum of just 5 species, including members of the 3 principal groups, to mimic the natural fermentation process and yield good quality chocolate. Reduction of the amount of pectin by physical or mechanical means can also lead t...

The Microbiology of Cocoa Fermentation and its Role in Chocolate Quality

When grown in the appropriate medium, several yeast species produce pectinases able to degrade pectic substances. It is mainly exocellular endopolygalacturonases that break pectins or pectate down by hydrolysis of α-1,4-glycosidic linkages in a random way. Biochemical characterisation of these enzymes has shown that they have an optimal pH in the acidic region and an optimal temperature between 40 and 55°C. Their production by yeasts is a constitutive feature and is repressed by the glucose concentration and aeration. Pectic substances and their hydrolysis products are used as carbon sources by a limited number of yeasts and hence these enzymes must be involved in the colonisation of different parts of plants, including fruits. The first yeast pectic enzyme (encoded by the PSE3 gene) was cloned from Tichosporon penicillatum. Recently, a polygalacturonase-encoding gene from Saccharomyces cerevisiae has been cloned and overexpressed in several strains and the gene for an extracellualar endopolygalacturonase from Kluyveromyces marxianus has also been described. Taking all the results together, the idea is now emerging that this type of yeast enzyme could offer an alternative to fungal enzymes for industrial applications.

Production of pectic enzymes in yeasts

Saccharomyces cerevisiae CECT1389 secreted an extracellular endopolygalacturonase (EC 3.2.1.15) when grown in shake flasks in medium containing galactose alone, or either galactose and polygalactur...

Production and partial characterization of an endopolygalacturonase from Saccharomyces cerevisiae

A structural polygalacturonase-encoding gene (PGU1) from Saccharomyces cerevisiae IM1-8b was cloned and sequenced. The predicted protein comprises 361 amino acids, with a signal peptide between residues 1 and 18 and two potential glycosylation points in residues 318 and 330. The putative active site is a conserved histidine in position 222. This polygalacturonase showed 54% homology with the fungal ones and only 24% homology with their plant and bacterial counterparts. The gene is present in a single gene copy per haploid genome and it is detected in all strains, regardless of their phenotype. The expression of PGU1 gene in several strains of S. cerevisiae revealed that the polygalacturonase activity depended on the plasmid used and also on the genetic background of each strain but in all cases the enzymatic activity increased.

Cloning, molecular characterization, and expression of an endo-polygalacturonase-encoding gene from Saccharomyces cerevisiae IM1-8b

Astaxanthin and other carotenoids from 29 mutant strains of the yeast Phaffia rhodozyma obtained by means of benomyl and/or ethyl-methanesulfonate treatment and extracted with dimethyl sulfoxide (DMSO) were separated by high-performance liquid chromatography. Detection at 474 nm revealed variations in the pigment content of the different mutant strains. Hypopigmented mutants showed higher β-carotene contents than the wild type, whereas hyperpigmented mutants exhibited considerable increases (up to 232%) in astaxanthin contents. Furthermore, and contrary to the wild type, the pigments in some of the mutants could be directly extracted with ethanol, and although the yield of pigment decreased in relation to DMSO, the content of astaxanthin increased up to 80%.

Analysis of astaxanthin and other carotenoids from several Phaffia rhodozyma mutants

BACKGROUND
Yeasts responsible for fermentation have an important repercussion on wine quality. This study presents the influence of two autochthonous strains of Saccharomyces cerevisiae (XG1 and XG3), a commercial yeast (QA23) and spontaneous fermentation on the chemical and sensory properties of wines from Godello and Albarino.

RESULTS
All the yeasts showed normal fermentative kinetics and were able to lead fermentations; therefore, they were responsible for wine chemical and sensory characteristics. Significant differences were found at the chemical level depending on yeast strain and variety. Albarino wines from XG1 and XG3 presented low total acidity and glycerol content. Godello wines from QA23 had higher total acidity but lower alcohol content than those from XG1, XG3 and spontaneous fermentation. QA23 wines presented a greater amount of higher alcohols and 2-phenylethanol for both grapevine cultivars, whereas XG3 and spontaneous fermentations yielded wines with a higher concentration of esters, mainly ethyl lactate, and fatty acids. These differences were detected at the sensory level; thus, Albarino made with XG3 and Godello from spontaneous fermentation were the most appreciated wines.

CONCLUSION
XG3, an autochthonous strain of S. cerevisiae, constitutes a useful tool to elaborate wines with singular characteristics. © 2013 Society of Chemical Industry

Pilar Blanco

Papers

Production of pectic enzymes in yeasts

Production and partial characterization of an endopolygalacturonase from Saccharomyces cerevisiae

Cloning, molecular characterization, and expression of an endo-polygalacturonase-encoding gene from Saccharomyces cerevisiae IM1-8b

Analysis of astaxanthin and other carotenoids from several Phaffia rhodozyma mutants

Fermentative aroma compounds and sensory profiles of Godello and Albariño wines as influenced by Saccharomyces cerevisiae yeast strains