Production of chitinolytic enzymes with Trichoderma longibrachiatum IMI 92027 in solid substrate fermentation

Thirty Trichoderma strains representing 15 species within the genus have been screened for extracellular production of chitinolytic enzymes in solid substrate fermentation (SSF). T.longibrachiatum IMI 92027 (=ATCC 36838) gave the highest yield (5.0 IU/g dry matter of substrate) after 3 days of fermentation on wheat bran-crude chitin (9:1 mixture) medium. The optimum moisture content (66.7%), chitin content (20%), initial pH of the medium (2-5) and time course (5 d) of SSF were determined for strain IMI 92027. No significant effect of different N and P additives was found on the chitinase yield in wheat bran-chitin mixture medium. Cellulase, xylanase, alpha-amylase and beta-xylosidase activities were also detected. The pH and temperature optima of chitinase complex of T.longibrachiatum IMI 92027 was found to be at 4.5 and 55 ℃, respectively. The enzyme totally lost its activity at 70 ℃ in 5 min in the absence of the substrate but retained about 15% of its initial activity even at 70 ℃ after incubation of 60 min in the presence of SSF solids (residual substrate and fungal mycelium). Purification of protein extract from the SSF material revealed high chitinolytic activities between pI 5.9.-4.8 where N-acetyl-β-D-hexosaminidase and chitinase peaks have been found in the same pI range. Chitinase peaks could be described at least at four pI values: pI 5.9; 5.6; 5.3 and 4.8 while at least four main N-acetyl-β-D-hexosaminidase peaks could be separated at pI 6.0 and 5.1 and at more basic isoelectric points of 7.2 and 8.0. Two chitinases with 43.5 kDa and 30 kDa were purified at acidic isoelectric point.

Xylanase production by Trichoderma strains in solid substrate fermentation

The importance of microbial enzymes in pulp and paper manufacturing has grown significantly in the last two decades. Solid substrate fermentation (SSF) holds tremendous potential for the production of microbial enzymes of commercial interest. SSF can be of special interest in those processes where the crude fermented product (whole SSF culture, in situ enzyme) may be used directly as the enzyme source. Xylanase preparations practically free of cellulase activity are especially useful for biobleaching of crude cellulose pulps. Thirty-nine Trichoderma isolates have been screened in SSF for xylanase production on hardwood oxygen-delignified soda-aq pulp as carbon source and enzyme inducer. Xylanase activities varied between 0 and 2200 IU/g dry matter (DM) of initial substrate. In most instances, the simultaneously produced cellulase levels were below 1.0 Filter Paper Unit (FPU) /g DM. The xylanase to cellulase activity ratio varied in the range of 5 to 3500. The three most promising isolates (TUB F-1647, TUB F-1658 and TUB F-1684) yielded xylanase activity of 2040, 1300 and 1500 IU/g DM xylanase, respectively, and 0.64, 0.43 and 0.43 FPU/g DM cellulase with a xylanase to cellulase activity ratio of 3200, 3000 and 3500, respectively. Wild strains F-1647, F-1658 and F-1684 were isolated from tree bark of Maldives, soils of Peru (last two), respectively. Medium optimization experiments to enhance the xylanase yield and to increase the xylanase to cellulase ratio have also been performed.

Possible link between congenital uterine malformations and epitheliod trophoblastic tumor

Background: Epitheloid trophoblast tumor (ETT) is a tenuous type of gestational trophoblastic disease. Although clinically ETT behave in variable fashion, it is considered as a potentially malignant tumor, fatal malignancy occurs in 10% of the cases. Case and Conclusion: A 41 years old patient, who previously underwent Tompkins operation because of congenital uterine septum was admitted to our clinic due to irregular bleeding. Despite of serum hCG levels under the upper limit of normal, histopathology analysis of D&C obtained endometrial samples confirmed the presence of ETT by immunostainings. Henceforth we performed prompt abdominal hysterectomy and preserved the normal structured ovaries. According to the müllerianosis theory of endometriosis aberrant differentiation or migration could cause spreading of cells throughout the fetal organogenesis of the uterus. These cells in an apoptosis week environment might correlate with the initiation of ETT. As a conclusion we suggest further studies to assess the link between ectopic cells and ETT, to gain a better understanding in the pathomechanism of the tumor.