Efficient Recovery of Lignocellulolytic Enzymes of Spent Mushroom Compost from Oyster Mushrooms, Pleurotus spp., and Potential Use in Dye Decolorization

This study was conducted in order to perform efficient extraction of lignocellulolytic enzymes amylase (EC 3.2.1.1), cellulase (EC 3.2.1.4), laccase (EC 1.10.3.2), and xylanase (EC 3.2.1.8) from spent mushroom compost (SMC) of Pleurotus ostreatus, P. eryngii, and P. cornucopiae. Optimal enzyme recovery was achieved when SMCs were extracted with 50 mM sodium citrate (pH 4.5) buffer at 4degrees C for 2 hr. Amylase, cellulase, and xylanase activities showed high values in extracts from P. ostreatus SMC, with 2.97 U/g, 1.67 U/g, and 91.56 U/g, respectively, whereas laccase activity and filter paper degradation ability were highest in extracts from P. eryngii SMC, with values of 9.01 U/g and 0.21 U/g, respectively. Enzymatic activities varied according to the SMCs released from different mushroom farms. The synthetic dyes remazol brilliant blue R and Congo red were decolorized completely by the SMC extract of P. eryngii within 120 min, and the decolorization ability of the extract was comparable to that of 0.3 U of commercial laccase. In addition, laccase activity of the SMC extract from P. eryngii was compared to that of commercial enzymes or its industrial application in decolorization.

Studies on in vitro degradability of mixed crude enzyme extracts produced from Pleurotus spp.

A preliminary investigation was conducted to assess lignocellulolytic efficiency of crude extracts from three white-rot fungi, Pleurotus florida PF05 (PF), Pleurotus sajor-caju PS07 (PS) and Pleurotus eryngii PE08 (PE). The activities of CMC-ase, xylanase, b-glucosidase, b-xylosidase, laccase and Mn peroxidase in extracts were evaluated. PF produced its highest CMC-ase (317 UL-1), b-glucosidase (62 UL-1), b-xylosidase (37 UL-1) and laccase (347 UL-1) activities while, PS produced highest xylanase (269 UL-1) and Mn peroxidase (69 UL-1) activities. In addition, crude extracts extracted were employed for their in vitro degradability assessment; and were evaluated with mono and mixed extracts separately to corn cob substrate. The losses in cell wall components and dry matter during 5 and 10 days incubations were analyzed after treatments of extracts. Maximum 8.2, 4.4 and 2.8% loss were found respectively in hemicellulose (HC), cellulose (C) and lignin (L) with mono extract of PF within 10 days. The influence of mono extract of each strain (PF, PS and PE) and their mixed extracts (PF+PS, PF+PE, PS+PE and PF+PS+PE) on degradation of cell wall constituents were remarkably differed. The mixed extract treatment proved maximum 13.6% HC loss by PF+PS+PE extract, 9.2% loss in C by PF+PS extract and 5.2% loss of L by the PF+PS+PE extract treatment. The highest dry matter loss (8.2%) was recorded with PF+PS+PE mixed extract combination.

Molecular and Morphological Characterization of Green Mold, Trichoderma spp. isolated from Oyster Mushrooms

Isolates of Trichoderma spp. collected from Pleurotus ostreatus and P. eryngii beds, which included loosened substrate compactness and development of green colour, were grouped into three species. The occurrence of different species of Trichoderma was as T. cf. virens (70.8%), T. longibrachiatum (16.7%) and T. harzianum (12.5%). The conidia of Trichoderma spp. were ellipsoidal, obovoid and phialides were bowling pins, lageniform and the length of phialides was 3.5~10.0 x 1.3~3.3 micromm. Phialides of T. cf. virens and T. harzianum were tending clustered, but it was solitary disposition in T. longibrachiatum. T. cf. virens was characterized by predominantly effuse conidiation, sparingly branched, and fertile to the apex and it was penicillate type. RAPD analysis could detect variability amongst three different species of Trichoderma using two newly designed URP-primers. However, intra-specific variation could not be detected in all the isolates except for rDNA sequence data classified Trichoderma isolates into three distinct groups representing three species. The profiles of rDNA sequences of isolates representing a species showed high similarity in T. cf. virens and T. harzianum. However, there was a variation in rDNA sequences of isolates representing T. longibrachiatum. The results of present study reveals that molecular techniques of RAPD and rDNA sequencing can greatly aid in classification based on morphology and precise identification of fast evolving species of Trichoderma.

Physiological Characteristics of Green Mold (Trichoderma spp.) Isolated from Oyster Mushroom (Pleurotus spp.)

This study was conducted to investigate physiological characteristics of Trichoderma spp. isolated from Pleurotus spp. Damage tests of Pleurotus spp. and mycotoxins tests of Trichoderma spp. were also done. The optimal growth temperature of Trichoderma spp. was 27~30degrees C. Although, T. longibrachiatum was able to grow at 37degrees C and grew 30~40 times faster than Pleurotus. The colony colour on PDA medium of T. cf. virens was yellowish green, T. longibrachiatum was yellow, and T. harzianum was turning to bright green. In damage tests of Pleurotus by Trichoderma, T. cf. virens caused the most severe damage to Pleurotus. T. longibrachiatum and T. harzianum caused less damage on Pleurotus but were able to cause greater damage to P. eryngii. One of the mushroom cultivars, P. ostreatus 8 was the most resistant to all Trichoderma spp.. Chitinolytic mycotoxin released by Trichoderma spp. caused 52.7% damage to Pleurotus. Mycotoxins released by T. longibrachiatum caused the greatest damaged (78.6%) on P. eryngii.