Marian Petre

Environmental Biotechnology for Bioconversion of Agricultural and Forestry Wastes into Nutritive Biomass

The cellulose is the most widely distributed skeletal polysaccharide and represents about 50% of the cell wall material of plants. Beside hemicellulose and lignin, cellulose is a major component of agricultural wastes and municipal residues. The cellulose and hemicellulose comprise the major part of all green plants and this is the main reason of using such terms as “cellulosic wastes” or simply “cellulosics” for those materials which are produced especially as agricultural crop residues, fruit and vegetable wastes from industrial processing, and other solid wastes from canned food and drinks industries.

Biotechnology of Agricultural Wastes Recycling Through Controlled Cultivation of Mushrooms

The agricultural wastes recycling with applications in agro-food industry is one of the biological challenging and technically demanding research in the biotechnology domain known to humankind so far. Annually, the accumulation of huge amounts of vineyard and winery wastes causes serious environmental damages nearby winemaking factories. Many of these ligno-cellulose wastes cause serious environmental pollution effects, if they are allowed to accumulate in the vineyards or much worse to be burned on the soil. At the same time, the cereal by-products coming from the cereal processing and bakery industry are produced in significant quantities all over the world (Moser, 1994; Verstraete & Top, 1992).

Controlled Cultivation of Mushrooms on Winery and Vineyard Wastes

Abstract The biotechnological processes for the total recovery of lignocellulosic wastes resulting from industrial processing of grapes, as well as the maintenance of vineyards, through in vitro cultivation of mushrooms were tested using three Basidiomycetes species, namely, Ganoderma lucidum (Curt.: Fr.) P. Karst, Lentinula edodes (Berkeley) Pegler, and Pleurotus ostreatus (Jacquin ex Fries) Kummer. The experiments in solid-state cultivation were done by in vitro growing of all these fungal species in special rooms, where the main culture parameters were kept at optimal levels in order to get the greatest production of mushroom fruit bodies. The effects of culture compost composition (carbon, nitrogen, and mineral sources) as well as other physical and chemical factors (such as temperature, inoculum amount, pH level, incubation time) on mycelial net formation, and especially on fruit body induction, were investigated. Also, a submerged cultivation of mushrooms for bioconversion of winery wastes into nutritive mycelia biomass was carried out to test the feasibility of such biotechnology.

Recycling of Vineyard and Winery Wastes as Nutritive Composts for Edible Mushroom Cultivation

Every year, in Romania huge amounts of wine and vine wastes cause serious environmental damages in vineyards as well as nearby winery factories, for instance, by their burning on the soil surface or their incorporation inside soil matrix. The optimal and efficient way to solve these problems is to recycle these biomass wastes as main ingredients in nutritive composts preparation that could be used for edible mushrooms cultivation. In this respect, the main aim of this work was to establish the best biotechnology of winery and vine wastes recycling by using them as appropriate growth substrata for edible and medicinal mushrooms. According to this purpose, two mushroom species of Basidiomycetes, namely Lentinula edodes as well as Pleurotus ostreatus were used as pure mushroom cultures in experiments. The experiments of inoculum preparation were set up under the following conditions: constant temperature, 23° C; agitation speed, 90–120 rev min−1 pH level, 5.0–6.0. All mycelia mushroom cultures were incubated...

Between two paradigms of mushroom biotechnology: solid-state cultivation and submerged growing

G arboreum is considered to be a rich source of stress responsive genes and EST data base revealed that most of its genes are uncharacterized. The full length Gossypium Universal Stress Protein-2 (GUSP-2) gene (510 bp) was cloned in Escherichia coli, Pichia pastoris and Gossypium hirsutum, characterized and point mutated at three positions separately at 352354, lysine-60 to proline (M1-usp-2) and 214-216, aspartic acid-26 to serine (M2-usp-2) and 145-147, lysine-3 to proline (M3usp-2) to study its role in abiotic stress tolerance. It was found that heterologous expression of one mutant (M1-usp-2) provided enhanced tolerance against salt and osmotic stresses, recombinant cells have higher growth up to 10-5dilution in spot assay as compared to W-usp-2 (wild type GUSP-2), M2-usp-2 and M3-usp-2 genes. M1-usp-2 in Pichia pastoris transcript profiling exhibited significant expression (7.1-fold) to salt and (9.7) and osmotic stresses. M1-usp-2 gene was also found to enhance drought tolerance and significant expression (8.7) in CIM-496-Gossypium hirsutum transgenic plants. However, little tolerance against heat and cold stresses both in recombinant yeast and bacterial cells was observed. The results from our study concluded that activity of GUSP-2 was enhanced in M1-usp-2 but wipe out in M2-usp-2 and M3-usp-2 response remained almost parallel to W-usp-2. Further, it was predicted through in silico analysis that M1-usp-2, W-usp-2 and M3-usp-2 may be directly involved in stress tolerance or function as signaling molecule to activate the stress adaptive mechanism.L oxidase (LOX) is an extracellular matrix, copper-dependent, amine oxidase that catalyzes a key crosslinking step in collagen and elastin. This enzyme has also been shown to play a role in promoting metastasis. The correlation between high LOX activity and cancer metastasis is strong enough that upregulated LOX activity can be used as a diagnostic marker for the severity of cancer in patients. β-aminopropionitrile is a known potent inhibitor of lysyl oxidase; however, this inhibitor is not selective and, therefore, cannot be used as a therapeutic agent. β-aminopropionitrile has been derivatized using aromatic sidechains and has been used to selectively target lysyl oxidase in breast cancer cells. The inhibitor LP-1-2 has been shown to reduce breast cancer cell viability with a 100 μM dose and 72-hour incubation period. The effect on cell viability increased with increasing amounts of inhibitor. The selective targeting of lysyl oxidase was verified using western blot analysis and lysyl oxidase activity assays. The activity assays showed that addition of increasing amounts of inhibitor decreased the activity of lysyl oxidase. The highest level of inhibition detected was with lysyl oxidase isolated from cells treated with 5000 μM of LP-1-2 for 3 days, which decreased the activity three-fold as compared to lysyl oxidase isolated from untreated cells.

Biotechnology of Mushroom Growth Through Submerged Cultivation

The submerged cultivation of mushrooms (SCM) requires full control of the bioprocess regarding the automatic tracking of all chemical and physical parameters, keeping them at optimal values. The main problem that needs to be solved by the intensive biotechnological process for submerged cultivation of edible and medicinal mushrooms on substrates made of agricultural wastes resulting from cereal grain processing is to convert these natural wastes of organic agriculture into nutritive biomass to be used as food supplements that are made through biological means only. The main stages of biotechnology to achieve highly nutritive mycelial biomass by controlled submerged fermentation are as follows: (i) preparation of culture substrates, (ii) steam sterilization of the bioreactor culture vessel, (iii) aseptic inoculation of sterilized culture media with the pure cultures of selected mushroom strains, (iv) running the submerged cultivation cycles under controlled conditions, and (v) collecting, washing, and filtering the fungal pellets which were obtained.

Biotechnological Recycling of Fruit Tree Wastes by Solid-State Cultivation of Mushrooms

Abstract The laboratory experiments which are presented in this chapter were carried out on testing and optimization of fruit tree wastes recycling through controlled cultivation of mushroom species Ganoderma lucidum and Pleurotus ostreatus in order to retrieve their carpophores to be used as food and nutritive supplements. To achieve these goals, a new and innovative environmental biotechnology was applied for full recovery and valorization of all fruit tree wastes (leaves, branches, dried trunks), usable as raw materials of hitherto untapped economic value, to prepare nutritive substrates for mushroom growth. In this way, the lignocellulosic wastes of fruit trees may be integrated extremely quickly into the main cycles of organic matter in nature, as new links introduced in the natural food chain by the cultivation of edible and medicinal mushrooms belonging to the mentioned species.