The hypersecreting mutant Trichoderma reesei RUT-C30 (ATCC 56765) is one of the most widely used strains of filamentous fungi for the production of cellulolytic enzymes and recombinant proteins, and for academic research. The strain was obtained after three rounds of random mutagenesis of the wild-type QM6a in a screening program focused on high cellulase production and catabolite derepression. Whereas RUT-C30 achieves outstanding levels of protein secretion and high cellulolytic activity in comparison to the wild-type QM6a, recombinant protein production has been less successful. Here, we bring together and discuss the results from biochemical-, microscopic-, genomic-, transcriptomic-, glycomic- and proteomic-based research on the RUT-C30 strain published over the last 30 years.

Trichoderma reesei RUT-C30--thirty years of strain improvement.

(1985). The Properties of Fungal and Bacterial Cellulases with Comment on their Production and Application. Biotechnology and Genetic Engineering Reviews: Vol. 3, No. 1, pp. 39-110.

https://www.tandfonline.com/doi/pdf/10.1080/02648725.1985.10647809

The Properties of Fungal and Bacterial Cellulases with Comment on their Production and Application

The global energy demand has been increasing rapidly due to depletion of fossil fuels, continuous growth of world population and industrialized economy. India has surpassed to Japan and Russia and become the third largest oil consumer in the world. Unfortunately, India’s primary energy consumption has increased due to reduced oil and gas production. The increased consumption of imported oil could lead to turbulence in economic growth. Due to increasing demand of oil fuels and consequent impact of global warming issues, development of alternate energy is a top priority in research and developments sector. The bioenergy produced from the biomass is being a sustainable alternate energy source which received high acceptance in various sectors include public, industries and government policies. from the Government, public, industries and researches for its sustainability. This review focuson bioprospecting of biomass from terrestrial and marine resources for non conventional energy production and the stepping stones of biofuel for near future. These carbohydrates can be converted into various forms of biofuels either directly or indirectly by exploiting microorganisms. However, the production process and chemical transformation is being an expensive process and therefore commercial supply of biofuel in largescale is not yet successful. Hence an economic and efficient production process is essential to commercialize biomass based biofuels. This article highlights the overview of sustainable and renewable resources for biofuel and stepping stones of biofuel commercialization.

Utilization of bioresources for sustainable biofuels: A Review

Worldwide photosynthetic fixation of carbon dioxide is estimated to yield annually up to 150 X 109 tons of dry plant material (biomass) (Lieth, 1973; Whittaker and Likens, 1973; Bassham, 1975; Stephens and Heichel, 1975). Almost half of this material consists of cellulose (28–50%); other major components are hemicelluloses (20–30%) and lignin (18–30%) (Thompson, 1983). Additional important but minor constituents of biomass are proteins, lipids, and carbohydrates such as chitin, starch, and pectin. A list of the amounts of cellulose in various plants has been compiled by Stephens and Heichel (1975). The biomass is eventually degraded and oxidized to CO2 and returned to the atmosphere. Thus, we have a carbon cycle. The gain of the cycling process is the capture of solar energy, which is then used by living cells for growth and maintenance.

Ecology of microbial cellulose degradation

Background
The discovery and development of novel plant cell wall degrading enzymes is a key step towards more efficient depolymerization of polysaccharides to fermentable sugars for the production of liquid transportation biofuels and other bioproducts. The industrial fungus Trichoderma reesei is known to be highly cellulolytic and is a major industrial microbial source for commercial cellulases, xylanases and other cell wall degrading enzymes. However, enzyme-prospecting research continues to identify opportunities to enhance the activity of T. reesei enzyme preparations by supplementing with enzymatic diversity from other microbes. The goal of this study was to evaluate the enzymatic potential of a broad range of plant pathogenic and non-pathogenic fungi for their ability to degrade plant biomass and isolated polysaccharides.

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Arsenal of plant cell wall degrading enzymes reflects host preference among plant pathogenic fungi

Increased endoplasmic reticulum content of a mutant of Trichoderma reesei (RUT-C30) in relation to cellulase synthesis

Two strains of Trichoderma reesei, wild type QM6a and mutant Rut-C30, were grown in meida containing an inducer, insoluble crystalline cellulose (Avicel PH101), as carbon source for 11 days. The cell growth, expressed as myceliar protein content, of Rut-C30 was 4–5 times higher than QM6a. The lack of ultrastructural disorganization, and absence of intracellular enzyme release into the growth medium, indicated that none of these two strains had undergone any significant autolysis during the entire growth phase. Cellulase activities, mainly endoglucanase, cellobiase and filter paper degrading activity (disc) were enhanced in Rut-C30 cells. A major change was observed in the endoglucanase activity which was 30 times higher in Rut-C30 than QM6a, whereas, both β-glucosidase and disc activities were 3 times enhanced in Rut-C30 compared to QM6a. In addition to synthesis, cellulase secretion was also enhanced in Rut-C30. Both the organisms contained same amounts of intracellular marker enzyme activities (e.g., inosine diphosphatase, thiamine pyrophosphatase, alkaline phosphatase). Finally, the enahncement of secretory activity of Rut-C30 was correlated with the proliferation of rough endoplasmic reticulum (RER) and increased phospholipid content. It appears that Rut-C30 is not only a hypercellulolytic but also a hypersecretor mutant.

Cellulase secretion from a hyper-cellulytic mutant of Trichoderma reesei Rur-C30

A growing trend in the development of viable processes for the conversion of renewable cellulosic biomass to glucose is the use of microbial cellulases as biological catalysts. Unfortunately, the cost of the cellulase enzymes has been prohibitive for large scale industrial application in saccharification of cellulose. The high cost of cellulase is due largely to the low yield and to the low specific activity of enzymes from the available microbial strains. Improvement of the cellulolytic microbial strains can be considerably enhanced through selective screening programs. However, the rationale for selection and the chance of isolation of more useful strains are hampered by our lack of understanding of mechanisms controlling the synthesis and secretion of cellulase. Successful genetic cloning and expression of the cellulase genes from Tviohodevma or any other cellulolytic microorganism is similarly dependent upon a sound basic knowledge of the control mechanisms. Tviohodevma genetics is an unexplored abyss. Although Tviohodevma is reported to have a sexual stage, in the Hypoovea (1,2) mating types are generally unavailable. Thus, traditional methods of delineating genetic linkages are elusive.

Regulatory Controls in Relation to Overproduction of Fungal Cellulases

Abstract Biorefineries are a model for greener production processes, based on the concept of bioeconomy. Instead of targeting first-generation biofuels—that compete with food supply—the focus relies on lignocellulosic material, considering many aspects, such as sustainable fuel production, as well as valorization of waste, as an alternative to the traditional petrochemical approach of goods production. Especially, in tropical countries agricultural activities lead to tremendous amounts of biomass, resulting in waste that has to be dealt with. In the case of Costa Rica, the five major crops cultivated for export are coffee, oil palm, pineapple, sugarcane, and banana. Traditional ways of waste treatment cannot cope with the increasing amount of biomass produced and therefore, bear various challenges often related to increased pollution. This review aims to bring up the recent state of waste treatment but even more, stress potential opportunities of adding value to not used residues; thus, improve sustainability in the agro industrial sector. Part I of the review already highlighted the potential of producing promising bioactive chemical compounds by novel biorefinery concepts from agricultural waste originating from coffee and oil palm cultivation. This second part focuses on the lignocellulose-rich biowaste from pineapple, sugarcane, and banana, showing biorefinery concepts, where fuel and energy production, as well as establishment of novel products and new applications, play an important role. 

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Tropical agroindustrial biowaste revalorization through integrative biorefineries—review part II: pineapple, sugarcane and banana by-products in Costa Rica

H. Trimiño-Vazquez

Papers

Increased endoplasmic reticulum content of a mutant of Trichoderma reesei (RUT-C30) in relation to cellulase synthesis

Cellulase secretion from a hyper-cellulytic mutant of Trichoderma reesei Rur-C30

Regulatory Controls in Relation to Overproduction of Fungal Cellulases

Tropical agroindustrial biowaste revalorization through integrative biorefineries—review part II: pineapple, sugarcane and banana by-products in Costa Rica