Colin G. Orpin

Bio: Colin G. Orpin is an academic researcher from Agricultural and Food Research Council. The author has contributed to research in topics: Rumen & Neocallimastix. The author has an hindex of 17, co-authored 20 publications receiving 1487 citations.

Studies on the rumen flagellate Neocallimastix frontalis.

Abstract: The vast increase in the population density of the rumen flagellate Neocallimastix frontalis shortly after the host animal has commenced eating is caused by stimulation of a reproductive body on a vegetative phase of the organism to differentiate and liberate the flagellates. The stimulant is a component of the host's diet. The vegetative stage of N. frontalis bears a strong morphological resemblance to that of certain species of aquatic phycomycete fungi, and consists of a reproductive body borne on a single, much branched rhizoid. The flagellates liberated in vivo within 15 to 45 min of feeding lose their motility within 1 h and develop into the vegetative phase, thus producing a rapid decrease in population density of the flagellates. Conditions for maximum flagellate production are similar to those ocfnring in the rumen: pH 6.5, 39 °C, absence of O2, presence of CO2. Differentiation of the reproductive body is inhibited by compounds affecting membrane structure and function, but not by inhibitors of protein synthesis. The organism was cultured in vitro in an undefined medium in the absence of bacteria or other flagellates.

Zoospore Chemotaxis in the Rumen Phycomycete Neocallimastix frontalis

Abstract: Neocallimastix frontalis zoospores showed chemotaxis to a range of carbohydrates, but not to the common amino acids, purines, pyrimidines or vitamins. Four chemoreceptors were identified: the glucose receptor, sensitive to D-glucose, D-galactose, D-xylose, L-sorbose, D-fucose and 2-deoxy-D-glucose; the sucrose receptor, sensitive to sucrose, D-fructose and raffinose; the mannose receptor, sensitive to D-mannose and D-glucose; and the sorbitol receptor, sensitive to D-sorbitol and D-mannitol. Growth of the vegetative stage of N. frontalis did not occur with D-xylose, L-sorbose, D-fucose, 2-deoxy-D-glucose, raffinose, D-mannose or D-sorbitol. The zoospores were attracted to carbohydrate mixtures representing the soluble carbohydrates found in different barley tissues; the highest response was with those mixtures representing carbohydrates of awn and inflorescence tissue. Chemotaxis also occurred preferentially to the awn and inflorescence tissue carbohydrate combinations rather than to carbohydrate combinations representing other tissues. Germination of the zoospores occurred in medium containing glucose in excess of 10-4 M.

The lipids of the rumen fungus Piromonas communis

Abstract: SUMMARY: The major phospholipids of the anaerobic rumen phycomycete Piromonas communis were phosphatidylethanolamine (38%), phosphatidylcholine (26%) and phosphatidylinositol (13%); no sphingolipids, glycolipids, plasmalogens or phosphonyl lipids were detected. Free fatty acids, triacylglycerols, 1:2 diacylglycerols and a variable amount of 1:3 diacylglycerol were identified, as were minor amounts of squalene and a triterpenol which is probably tetrahymanol. Approximately half the fatty acids were straight chain, even 12 to 24 carbon, saturated acids, the remainder being even 16 to 24 carbon, mono-unsaturated fatty acids. The double bonds in all except the 16 carbon acid were in the ω9 position. The unsaturation is introduced by a δ9 desaturase which uses stearic acid as substrate and which does not use oxygen as a terminal electron acceptor. 14C from acetate and glucose was incorporated into the fatty acids of all complex lipids, as were lauric, myristic, palmitic, stearic and oleic acids. [14C]Choline was incorporated into phosphatidylcholine and [14C]ethanolamine into phosphatidylethanolamine and phosphatidylcholine. Label from [14C]serine was recovered in phosphatidylserine and phosphatidylethanolamine, but was not detected in phosphatidylcholine.

Microbial cellulose utilization: fundamentals and biotechnology.

Abstract: 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.

The Rumen Protozoa

Abstract: In addition to the bacteria in the rumen there are many larger (5–250/mm long) organisms which at various times have been designated protozoa Of these the ‘ovals’ (Quin’s and Eadie’s) are now known to be large bacteria (Orpin, 1976) and the ‘flagellates’ Neocallimastix frontalis, Piromonas communis and Sphaeromonas communis are the zoospores of phycomycete fungi (Orpin, 1977a, b) There are genuine flagellates in the rumen, eg Trichomonas spp, Monoceromonas sp and Chilomastix sp, but almost nothing is known about their metabolism (Jensen and Hammon, 1964) The largest, most obvious and most important protozoa are the ciliates, of which there are two groups both in the subclass Trichostomatia The so called ‘holotrich’ protozoa belong to the order Vestibuliferida and the entodiniomorphs to the order Entodiniomorphida, suborder Entodiniomorphina and family Ophryoscolecidae As the properties and metabolism of these two protozoal groups are different, they will be considered separately below

Fungal bioconversion of lignocellulosic residues; opportunities & perspectives.

Abstract: The development of alternative energy technology is critically important because of the rising prices of crude oil, security issues regarding the oil supply, and environmental issues such as global warming and air pollution. Bioconversion of biomass has significant advantages over other alternative energy strategies because biomass is the most abundant and also the most renewable biomaterial on our planet. Bioconversion of lignocellulosic residues is initiated primarily by microorganisms such as fungi and bacteria which are capable of degrading lignocellulolytic materials. Fungi such as Trichoderma reesei and Aspergillus niger produce large amounts of extracellular cellulolytic enzymes, whereas bacterial and a few anaerobic fungal strains mostly produce cellulolytic enzymes in a complex called cellulosome, which is associated with the cell wall. In filamentous fungi, cellulolytic enzymes including endoglucanases, cellobiohydrolases (exoglucanases) and β-glucosidases work efficiently on cellulolytic residues in a synergistic manner. In addition to cellulolytic/hemicellulolytic activities, higher fungi such as basidiomycetes (e.g. Phanerochaete chrysosporium) have unique oxidative systems which together with ligninolytic enzymes are responsible for lignocellulose degradation. This review gives an overview of different fungal lignocellulolytic enzymatic systems including extracellular and cellulosome-associated in aerobic and anaerobic fungi, respectively. In addition, oxidative lignocellulose-degradation mechanisms of higher fungi are discussed. Moreover, this paper reviews the current status of the technology for bioconversion of biomass by fungi, with focus on mutagenesis, co-culturing and heterologous gene expression attempts to improve fungal lignocellulolytic activities to create robust fungal strains.

Cellulose hydrolysis by bacteria and fungi

Abstract: Publisher Summary The chapter focuses on the recent advances in understanding the structural and functional organization of individual cellulases, their regulation, and the ways in which the multiple enzyme components of cellulolytic systems cooperate. It overviews the cellulose structures because cellulose is more than a homopolymer of β-1 ,4 linked glucose units. An appreciation of its complex physical organization and its interactions with other plant cell wall components is central for understanding of the mechanisms of cellulase action. Cellulose nearly always occurs in close association with plant cell wall matrix polysaccharides so that enzymes such as xylanases are intimately involved in the attack of cellulose in vivo. Cellulases effect important changes to their substrate before releasing soluble products and the key to understanding cellulase action rest in the examination of these events. Progress in this area is limited by the availability of appropriate analytical tools although new techniques, such as atomic force microscopy are promising. The properties of cellulases are profoundly altered by the presence of trace enzyme contaminants. Future studies in vitro are proposed to be restricted to enzymes from recombinant sources. The reasons for the individual cellulolytic bacteria and fungi requiring many related cellulases with specificities that overlap is still not clear, but perhaps this is because the complexity of the substrates and of the task these microorganisms face is underestimated.