Production and structural features of a water-soluble polysaccharide from a mutant strain of Agrobacterium sp.

We have developed a mutant strain derived from Agrobacterium sp. ATCC 31750, which produces a water-soluble polysaccharide having potential utility to the food, feed, pharmaceutical and cosmetic industries. A high concentration of product (15 g/L) is obtained by 48 h cultivation of the mutant strain under optimized fermentation conditions. The water-soluble polysaccharide obtained from cultures of the mutant strain beta82 has Glc:Man:Gal in approximate molar ratios of 5.8:6.7:1.0. The molecular weight of the polysaccharide was determined to be approximately 1000 kDa by HPSEC analysis. Linkage analysis contained 3-Glcp, 3-Manp, terminal Glcp and terminal Manp, as well as a small proportion of 3- and 3,4-Galp, and 4,6-Manp residues. Based on analyses using FT-IR and ¹³C NMR spectrometers, most glycosidic bonds joining these sugar residues are of the α-type, and acetyl groups are apparently attached to the polymer chain at random.     

Microbiological safety of kinema, a fermented soya bean food

Kinema is a fermented soya bean food of Nepal and the hilly regions of North-eastern States of India. Generally, the fermentation is dominated by Bacillus spp. that often cause alkalinity and desirable stickiness in the product. The present study was undertaken in a limited number of commercial (market) kinema samples to test for the presence of foodborne pathogens and their properties. Bacillus cereus was present in numbers exceeding 10⁴ cfu/g product in five of the tested 15 market samples. Enterobacteriaceae and coliform bacteria exceeded 10⁵ cfu/g in 10 of the 15 samples. Escherichia coli exceeding 10⁵ cfu/g was found in two samples. Staphylococcus aureus was not detected in any of the tested samples. Of 31 isolated typical and atypical strains of B. cereus, 18 representative strains were tested qualitatively for the ability to produce diarrhoeal type enterotoxin (BCET) using an Oxoid BCET-RPLA test kit. Overall, BCET was formed by 12 strains in BHIG (brain heart infusion broth +1% glucose), by seven strains on sterilized cooked rice, and by five strains on sterilized cooked soya beans. Semi-quantitative tests on BCET revealed that levels exceeding 256 ng/g soya beans, produced by single pure culture inoculation with the isolated B. cereus strains, were reduced to ≤ 8 ng/g by frying kinema in oil, a common procedure when making kinema curry. It was also shown in a mixed pure culture experiment that a kinema strain B. Subtilis DK-W1, is able to suppress growth and BCET formation by a selected toxin producing strain (BC7-5) of B. cereus. It is concluded that the traditional way of making kinema and its culinary use in curries is safe. However, for novel applications of kinema, safety precautions are advisable.     

The Effect of Ozone Gas-Liquid Contacting Conditions in a Static Mixer on Microorganism Reduction

The effect of gas-liquid contacting conditions in a static mixer on ozone transfer efficiency and reduction of Bacillus subtilis spores was studied in an experimental ozone contactor. An empirical mathematical model was developed that related the transfer efficiency in the experimental system to the superficial liquid velocity in the mixer, the gas-liquid flow rate ratio and the height of the down-stream bubble column. Spore reduction was determined primarily by the dissolved ozone concentration-time (C_avgt_m) product in the reactive flow segment and was independent of the gas-liquid contacting conditions in the static mixer. In an integrated ozone contacting system, the static mixer should be designed to maximize ozone mass transfer while the reactive flow segment should be designed for efficient microorganism reduction.     

Comparative study of the elimination of toluene vapours in twin biotrickling filters using two microorganisms Bacillus cereus S1 and S2

BACKGROUND: To investigate the microbial degradation performance of organic pollutants in the atmosphere using a biotrickling filter, two microorganism strains, Bacillus cereus S1 and Bacillus cereus S2, were selected, identified and inoculated into a twin biotrickling filter for comparison. RESULTS: Both strains showed good performance towards the degradation of model organic pollutants when gas flow rates ranged from 100 to 600 L h⁻¹. For S1, the total maximum removal efficiency (RE) of toluene was maintained nearly 100% not only at gas flow rates of 100 L h⁻¹ corresponding to empty bed residence time (EBRT) 199.44 s, but also at gas flow rates of 200 L h⁻¹ (EBRT = 99.72 s) and 300 L h⁻¹ (EBRT = 66.48 s). However, S2 had a much lower degradation capability; near 100% removal efficiency was obtained only at the gas flow rate of 100 L h⁻¹ although both bacteria belong to the same Bacillus cereus. With further increase in gas flow rate, the total REs for both S1 and S2 decreased slightly at first and then dropped sharply to 46% and 35%, respectively, at an EBRT of 33.24 s, corresponding to a gas flow rate of 600 L h⁻¹. Starvation for between 2 and 10 days resulted in the re‐acclimation times of both strains ranging between 1.0 and 15.5 h. CONCLUSION: Strain S1 would be a better choice for inoculation into a biotrickling filter than strain S2, because of its much higher toluene removal capacity and rapid recovery to full performance. Copyright © 2008 Society of Chemical Industry     

Accelerator Analysis of Tributyltin Adsorbed onto the Surface of a Tributyltin Resistant Marine Pseudoalteromonas sp. Cell

Tributyltin (TBT) released into seawater from ship hulls is a stable marine pollutant and obviously remains in marine environments. We isolated a TBT resistant marine Pseudoalteromonas sp. TBT1 from sediment of a ship’s ballast water. The isolate (10^{9.3 ± 0.2} colony-forming units mL⁻¹) adsorbed TBT in proportion to the concentrations of TBTCl externally added up to 3 mM, where the number of TBT adsorbed by a single cell was estimated to be 10^8.2. The value was reduced to about one-fifth when the lysozyme-treated cells were used. The surface of ethanol treated cells became rough, but the capacity of TBT adsorption was the same as that for native cells. These results indicate that the function of the cell surface, rather than that structure, plays an important role to the adsorption of TBT. The adsorption state of TBT seems to be multi-layer when the number of more than 10^6.8 TBT molecules is adsorbed by a single cell.     

胨冻样芽孢杆菌(Bacillus mucilaginosus)对烟草灰霉病菌(Botrytis cinerea)的抑制作用

本文首次报道胨冻样芽孢杆菌XDB1和D4B1对烟草灰霉病菌(Botrytis cinerea)的抑制作用。实验表明,XDB1和D4B1的发酵原液和无细胞提取物具有相同的抑菌效果,而经121℃处理30分钟后,抑菌活性丧失。D481菌株的抑菌活性高于XDB1,钾长石作为基质时的活性高于土壤矿物。但是,XDB1和D481对立枯丝核病菌(R..solani)、西瓜尖孢镰刀菌(F.oxysporum)、腐霉(Pythium sp)、小麦全蚀病菌(G.graminis)、烟炭疽病菌(C.gloeosporides)、烟草赤星病菌(A.alternata)、烟草黑胫病菌(P.nicotianae、串珠镰刀病菌(F.moniliforme)和水稻稻瘟病菌(P.grisea)无抑制作用。     

Surface tension as a limiting factor for aerobic n-alkane biodegradation

A new mathematical model for n-alkane biodegradation in crude oil, heavy oil and paraffinic mixtures is described. The pattern of n-alkane degradation as a function of the inverse of hydrocarbon chain length reported in this paper can be considered as general behaviour for many aerobic n-alkane biodegradation processes. A new interpretation of n-alkane biodegradation as a function of surface tension, is given. A mathematical expression was obtained starting from the degradation values of n-alkane and relative surface tension, which is a parameter independent of fermentation conditions. An interesting parameter, b, was identified which represented the accelerating conversion factor for n-alkane biodegradation. The findings suggested that the n-alkane biodegradation. The findings suggested that he n-alkane biodegradation rate may be affected by the fermentation condition (agitation, aeration, etc.) and by the strain of microorganism, while the behaviour pattern of n-alkane degradation was essentially linked to the substrate characteristics (molecular structure, molecular weight and density).     

枯草杆菌碱性磷酸酶制备的最佳工艺条件及酶促反应动力学性质的研究

以枯草杆菌为菌种液体培养制备碱性磷酸酶，研究确定了培养的最佳工艺条件，并对碱性磷酸酶的酶促反应动力学性质进行了初步探讨。结果表明，枯草杆菌制备碱性磷酸酶的最佳工艺条件为：40℃，pH值7．4振荡培养10h，酶活最高。对碱性磷酸酶的动力学性质研究表明，该酶催化底物磷酸苯二钠水解反应的最适pH值8．8，最适温度52℃，米氏常数Km值为2．94mmol／L。     

Hydrolysis of human horny cells by alkaline protease: Morphological observation of the process

The interaction of highly purified alkaline protease fromBacillus sp. KSM-K16 with the horny cells of human skin contained in skin grime was directly visualized by electron microscopy. It became clear that the protease first penetrates the horny cells and then adsorbs, mainly onto the internal structure of the cells at the initial stage of hydrolysis, and directly hydrolyzes the keratin filaments, though the marginal band surrounding them retains its original shape. Then, hydrolysate produced from the keratin filaments flows out of the cell, and early in the hydrolysis process keratin filaments decrease and then disappear, leaving a marginal band, i.e., the cell turns to a hollow state. As a result, the remaining marginal band loses support from inside the cell, thus promoting cleavage and dispersion. Until this stage in the protease reaction, the remarkable liberation of hydrolysis products as water-soluble protein does not occur.     

Monooxygenase system of <Emphasis Type="Italic">Bacillus megaterium</Emphasis> ALA2: Studies on palmitic acid oxidation products

We identified many novel oxygenated FA produced from linoleic acid by microbial strain ALA2: 12,13,17-trihydroxy-9(Z)-octadecenoic acid (12,13,17-THOA); 12,13,16-trihydroxy-9(Z)-octadecenoic acid (12,13,16-THOA); 12-hydroxy-13,16-epoxy-9(Z)-octadecenoic acid; and 12,17;13,17-diepoxy-16-hydroxy-9(Z)-octadecenoic acid. 12,13,17-THOA, the main product, inhibits the growth of some plant pathogenic fungi. Recently, we reclassified strain ALA2 as Bacillus megaterium ALA2 NRRL B-21660 and opened a possible link with the well-studied catalytically self-sufficient P450 monooxygenase of Bacillus megaterium ATCC 14581 (NRRL B-3712) and B. subtilis strain 168 (NRRL B-4219). Now we have found that strain ALA2 also oxidizes palmitic acid into three oxygenated products: 13-, 14-, and 15-hydroxy palmitic acids. This indicates that strain ALA2 also possesses a monooxygenase system similar to the abovementioned well-known strains. These data facilitate studies on the oxygenase system of strain ALA2.