Selection of antibiotic-resistant mutants with enhanced isoamylase activity in Pseudomonas amyloderamosa

Summary Isoamylase-hyperproducing strains of Pseudomonas amyloderamosa were bred by mutagenesis with UV light and N-methyl-N-nitro-N-nitrosoguanidine (NTG). The selection criterion for such strains was based on the formation of large turbid zones around the bacterial colonies in agar medium containing antibiotics and 1% waxy corn starch. Mutant WN6410 was obtained by treating P. amyloderamosa JD210 with five cycles of 1 × 10⁴ J UV light and one cycle of NTG. P. amyloderamosa WN6410 had 22-fold increase in isoamylase activity when compared to wild-type strain SB15 and the maximal enzyme activity, 5,100 U/ml, could be achieved within 48 h in 2.5 L fed-batch fermentation.     

Screening and production of erythritol by newly isolated osmophilic yeast-like fungi

Twenty-eight erythritol-producing strains were isolated from pollen, honey and high sugar food samples collected in Taiwan. Amongst these, six strains (166-2, 262-1, 278-3, 440, 441 and 442) were high erythritol-producers with a yield higher than 30% for 30% glucose. The erythritol productivity of these strains ranged from 90.9 to 116.4 g l⁻¹. ¹H- and ¹³C-NMR analyses confirmed that the fermentation product was erythritol. The results of morphological and physiological studies indicate that strains 166-2, 262-1, 278-3, 440, and 442 may be members of the genus Moniliella. More studies are required to determine the taxonomic position of strain 441. The use of a medium containing 30% glucose and 1% yeast extract gave the highest erythritol productivity. On batch fermentation in a 5-l fermentor using strain 166-2, a maximal erythritol productivity of 111.0 g l⁻¹ was obtained after cultivation for 144 h.     

Variation in rare earth element (REE), aluminium (Al) and silicon (Si) accumulation among populations of the hyperaccumulator Dicranopteris linearis in southern China

Plant and Soil - Dicranopteris linearis is a rare earth element (REE), aluminium (Al) and silicon (Si) hyperaccumulator plant which occurs in southern China. To date, there have been no studies on...     

Effect of pH on isoamylase production by Pseudomonas amyloderamosa WU 5315

The isoamylase activity of Pseudomonas amyloderamosa WU 5315 was stable over the pH range from 5.5 to 6.25 while only about 30% of the activity remained at pH 6.5. Low isoamylase activity (418 U ml^-1) was produced by the cells grown at high pH. Activity reached almost 3000 U ml^-1 when pH was kept below 6.0 during the fermentation. With 1% glucose plus 2% maltose instead of 3% maltose as carbon source, however, no pH control was required and the isoamylase activity of Ps. amyloderamosa WU 5315 increased to 3400 U ml^-1.     

Indicator species drive the key ecological functions of microbiota in a river impacted by acid mine drainage generated by rare earth elements mining in South China

Acid mine drainage (AMD) generated by rare earth elements (REEs) deposits exploration contains high concentrations of REEs, ammonium and sulfates, which is quite different from typical metallic AMD. Currently, microbial responses and ecological functions in REEs-AMD impacted rivers are unknown. Here, 16S rRNA analysis and genome-resolved metagenomics were performed on microbial community collected from a REEs-AMD contaminated river. The results showed that REEs-AMD significantly changed river microbial diversity and shaped unique indicator species (e.g. Thaumarchaeota, Methylophilales, Rhodospirillales and Burkholderiales). The main environmental factors regulating community were pH, ammonium and REEs, among which high concentration of REEs increased REEs-dependent enzyme-encoding genes (XoxF and ExaF/PedH). Additionally, we reconstructed 566 metagenome-assembled genomes covering 70.4% of identifying indicators. Genome-centric analysis revealed that the abundant archaea Thaumarchaeota and Xanthomonadaceae were often involved in nitrification and denitrification, while family Burkholderiaceae were capable of sulfide oxidation coupled with dissimilatory nitrate reduction to ammonium. These indicators play crucial roles in nitrogen and sulfur cycling as well as REEs immobilization in REEs-AMD contaminated rivers. This study confirmed the potential dual effect of REEs on microbial community at the functional gene level. Our investigation on the ecological roles of indicators further provided new insights for the development of REEs-AMD bioremediation.     

Efficient purification of transglutaminase from recombinant <Emphasis Type="Italic">Streptomyces platensis</Emphasis> at various scales

An efficient system for the fast and efficient purification of transglutaminase from recombinant Streptomyces platensis and expressed in Streptomyces lividans 25-2 is described. Because the purification procedure of this system is flexible, culture broth from laboratory (20 l) and pilot-plant (130 l) fermentations were used to purify the enzyme to electrophoretic homogeneity with high purity (90–95%) and yield (61–77%) within 1 or 2 days.     

昆虫细胞生长之调控机制的探讨

细胞生长调控机制的探讨是近年来发育生物学中一个十分热门的研究领域。生物体内之细胞是如何得知何时该生长及分裂?何时该停止生长?何时该死亡?对生物体来说至关重大。研究显示调控细胞生长之信息传递系统出现差错,将导致生长异常,从而产生组织细胞之异常增生而诱发癌症或产生其它重大疾病。而人类也只有在充分理解细胞生长之机制的基础上,我们才能了解癌症等重大疾病发生的细胞生物学基础。在探讨细胞生长调控机制的研究中,昆虫特别是果蝇Drosophilamelanogaster一直是一个十分理想的实验材料。文章介绍了如何从昆虫看细胞之生长调控机制。     

Screening, purification, and characterization of an extracellular prolyl oligopeptidase from Coprinopsis clastophylla

Culture filtrates of 22 mushrooms were screened for extracellular prolyl oligopeptidase activity. Four strains with relatively high enzyme activity were all from inky cap mushrooms. The production of Coprinopsis clastophylla prolyl oligopeptidase was associated with the growth of the fungus and the enzyme was not released by cell lysis. The enzyme was purified 285-fold to a specific activity of 52.05 U/mg. It was purified to a single band on a native polyacrylamide gel. However, the enzyme separated into three bands on a sodium dodecyl sulfate-polyacrylamide gel with mobility corresponding to molecular weights of approximately 84, 60, and 26 kDa. The results of tandem mass spectrometric analysis revealed that the 60 kDa protein was likely a degradation product of the 84 kDa protein. The isoelectric point of the purified enzyme was 5.2. The purified enzyme had an optimal pH and temperature of 8.0 and 37°C, respectively. Diisopropyl fluorophosphate (DFP), p-chloromercuribenzoaic acid (PCMB), Hg(2+), and Cu(2+) strongly inhibited C. clastophylla prolyl oligopeptidase. This enzyme is a serine peptidase and one or more cysteine residues of the enzyme are close to the active site.     

Different types of variant muscle nociception after intermittent and continuous neuromuscular stimulation in rats

Summary Critical assessment of experimental muscle-pain models resulting from maximal muscle contraction may provide a means of assessing hypersensitivity and the central nociceptive mechanisms involved in diffused muscle pain. The aim of the present study was, therefore, to investigate the patterns of nociceptive behavior and neuronal changes in the rat spinal cord after two modes of maximal muscle contraction. The gastrocnemius muscle of adult male Sprague-Dawley rats was subjected to continuous (10 min) or intermittent (60 min, 10/50 s on/off ratio) premodulated electrical stimulation of median frequency. Similar peak forces but different patterns of contraction output were generated by these two stimulation modes. Nociceptive behavioral scores and hind-leg oedema were significantly greater in the continuous group compared to the controls; however, significant difference was not demonstrated for either parameter comparing the intermittent and control groups. The sensory threshold was slightly reduced after the intermittent stimulation, and elevated after the continuous modality. The elevation of sensory threshold could be reversed by naloxone administration. More Fos-labeled nuclei were noted for both of the stimulation groups relative to the controls. The Fos-labeled nuclei were larger for the intermittent group than for the continuous and control analogs. The results of the present study suggest that prolonged contraction from continuous stimulation results in specific nociceptive neuron activation, muscle lesion and endogenous opioid release causing exaggerated nociception.     

Bioconversion of cephalosporin C with D-amino acid oxidase from the yeastRhodosporidium toruloides

Summary A D-amino acid oxidase-producing yeast,Rhodosporidium toruloides CCRC 20306, was used to convert cephalosporin C (Ceph C) into -ketoadipyl cephalosporanic acid. A part of Ceph C could be directly converted into glutaryl-7-aminocephalosporanic acid (GL-7-ACA) by permeated cells of CCRC 20306. There were unknown side products formed during the conversion. The side products could be substantially reduced in amount by heating the cell extract containing D-alanine at 55°C for 5 min for five cycles prior to the conversion. Esterase activity present in the cell extract, which might be one of the causes of the side reaction, was greatly diminished by heat treatment.