Somatic cell fusion of Trichoderma reesei resulting in new genetic combinations

Summary A selection method has been developed for the isolation of recombinant strains of Trichoderma reesei QM 9414. The method is based on somatic hybridization via anastomosis or protoplast fusion, and on the difference in growth rate of the resulting heterokaryons and synkaryons. The more intensive growth of the synkaryons as due to allelic complementation of adenine-requiring auxotrophic strains mutated in the adenylosuccinate synthetase gene. The synkaryons appeared is energetically growing spots in the heterokaryotic background. Stable diploids could not be isolated, which points to the transient nature of the diploid state in this species.     

Purification and characterization of the catalytic subunit of cAMP dependent protein kinase from Drosophila melanogaster

Polyethylenimine selectivity precipitates a large fraction of the proteins present in a crude Drosophila embryonic extract. While the free catalytic subunit of the cAMP dependent protein kinase is quantitatively retained in the soluble fraction after polyethylenimine precipitation, the rest of the abundant and highly active protein kinases present in the embryo are quantitatively precipitated. The catalytic subunit of the cAMP dependent protein kinase was purified until apparent homogeneity from the soluble protein fraction after polyethylenimine precipitation. The pH optimum of the purified enzyme is 6.3. While magnesium is the preferred divalent cation for all the cAMP dependent protein kinases described previously, the Drosophila enzyme is three times more active if manganese is present as divalent cation compared with magnesium. The enzyme is tost active between 50–100 mM monovalent ion concentration. Heparin can selectively modulate the phosphorylation of different substrate proteins.     

Endonuclease-free, protoplast-forming enzyme preparation and its application in fungal transformation.

An endonuclease-free, protoplast-forming enzyme complex was prepared from the "snail enzyme." The purified preparation has high protoplast-forming activity comparable to the crude enzyme complex without destroying circular plasmid DNA. Furthermore, a higher transformation rate was achieved by the application of the endonuclease-free enzyme complex in both yeast and filamentous fungal vector-host systems.     

Proton magnetic resonance studies of bradykinin antagonists

Bradykinin (BK) is a peptide hormone with sequence Arg¹-Pro²-Pro³-Gly⁴-Phe⁵-Ser⁶-Pro⁷-Phe⁸-Arg⁹ and has been implicated in a multitude of pathophysiological processes such as the ability to lower systemic blood pressure and stimulate pain. BK analogues having bulky, β-branched D -aliphatic residues at position 7 combined with bulky L -aliphatic residues at position 8 have now been observed to be strong antagonists. Conformational studies based on two-dimensional nmr experiments in methanol/water (80/20 v/v) were carried out on several such active antagonists in a polar solvent. Included in this study were the very active antagonists, [D -Arg⁰, Hyp³, Thi⁵, D -Cpg⁷, Cpg⁸]-BK [Cpg: α-cyclo-pentyl-glycine; Hyp: trans-4-hydroxy-L -proline; Thi: β-(2-thienyl)-L -alanine] ( I ), [D -Arg⁰, Hyp³, D -Cpg⁷, Cpg⁸] -BK ( II ), as well as its variant with D -Cpg⁷ replaced by Cpg⁷, namely [D -Arg⁰, Hyp³, Cpg⁷, Cpg⁸]-BK ( III ). A turn-like structure, which coexists with the extended conformation, was observed between residues 2 and 5 for the most active antagonists I and II , in direct correlation with the peptide activities. No turn-like structure was found for residues 6–9. In peptide III , a turn-like structure was not identified. The existence of a turn at the C-terminal end of bradykinin and its analogues has been predicted by empirical calculations and supported by nmr measurements. But the present nmr study on the most active antagonists ( I , II ) does not support this hypothesis. Instead, the data suggest that a turn-like structure between residues 2 and 5 could be important for antagonist activity. Finally, one weak inhibitor [D -Cpg⁷]-BK ( IV ) showed no defined secondary structure. © 1993 John Wiley & Sons, Inc.     

A new mutation protocol for obtaining auxotrophic mutants of the yeastPhaffia rhodozyma

Summary A new mutation strategy, which involves -irradiation of cells followed by a selective enzymatic enrichment step, was worked out to obtain auxotrophic mutants from the astaxanthin-producing yeast P. rhodozyma. Under the optimized conditions described, different mutants suitable for strain improvement were isolated.     

Teucrins H1-H4, novel clerodane-type diterpenes from Teucrium hyrcanicum

The structure and stereochemistry of teucrins H1, H2, H3 and H4, four novel diterpenes from Teucrium hyrcanicum have been established by detailed studies of the PMR and ¹³C NMR spectra, CD results and chemical transformations. Teucrin H4, a minor constituent is shown to possess a unique stereochemistry hitherto not reported for related diterpenoids.     

Phytoplankton biomass and chlorophyll-a in some shallow lakes in central Europe

Based on 388 parallel data of phytoplankton biomass and chlorophyll-a of two shallow lakes and two ponds, the following results were obtained:

1. Relative chlorophyll-a content of phytoplankton biomass varied between 0.08–1.88%; chlorophyll-a concentration showed great differences among lakes.
2. Significant correlations (r = 0.68–0.92) were established between phytoplankton biomass and chlorophyll-a concentration. The regression in the artificial ponds was non-linear.
3. In parallel with the increase of average cell volume, a decrease in relative chlorophyll-a content was observed. A significant correlation (r = + 0.83) between the two variables was found. Relative chlorophyll-a content of phytoplankton is a log-function of average cell volume.