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Query: UMLS:C0040822 (tremor)
 18,428 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

To examine the secretory production of heterologous proteins by Streptomyces lividans, we fused the DNA encoding the signal peptide of the alpha-amylase inhibitor tendamistat, derived from S. tendae with a synthetic gene encoding the thrombin inhibitor hirudin. The analysis of secretion by immunoblots revealed an efficient translocation of hirudin through the membrane, with no detectable immunoreaction among the cellular proteins. The secreted hirudin was stable in the shaking culture for about 6 days. A comparison of the hirudin secreted by S. lividans and recombinant reference hirudin from yeast by immunoblots and thrombin inhibition assays shows that hirudin from Streptomyces has a lower specific activity, which may be due to a different aminoterminal sequence or to inexact processing of the precursor.
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PMID:Synthesis and secretion of hirudin by Streptomyces lividans. 136 34

The paper describes shake flask culture optimization studies for the production of amylases by Thermomyces lanuginosus. The culture was found to produce maximally alpha-amylase and glucoamylase (18.4 & 11.2 units/ml), respectively when the medium contained rice flour (2% w/v) as carbon and corn steep liquor as nitrogen source with medium pH adjusted to 5.5 and incubated for 72 h under shaking conditions (150 rpm) at 50 degrees C.
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PMID:Shake culture studies for the production of amylases by Thermomyces lanuginosus. 933 Jun 67

An external-loop airlift bioreactor, with a low ratio 2.9 of height-to-diameter of the riser and a ratio 6.6 of riser-to-downcomer diameter, was used to produce alpha-amylase from fermentation with dregs by Bacillus subtilis. The effects of gas flow rate and liquid volume on alpha-amylase production were investigated. After a 36-h fermentation time, an average of 432.3U/ml alpha-amylase activity was obtained under the conditions of liquid volume 8.5l and gas flow rate 1.2vvm for the first 12h of fermentation, 1.4vvm from 12 to 27h, and 1.2vvm from 27h to the end. The activity was higher than that obtained in shaking flasks (409.0U/ml) and in a mechanically stirred tank bioreactor (397.2U/ml) under optimized operating conditions. The fermentation cycle of the airlift bioreactor was shorter than the 48h required for the shaking flasks and close to the 36h of the mechanically stirred tank bioreactor. It was demonstrated that the external-loop airlift bioreactor could substitute for the traditional mechanically stirred tank bioreactor to produce alpha-amylase from fermentation by Bacillus subtilis with dregs.
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PMID:alpha-Amylase production by Bacillus subtilis with dregs in an external-loop airlift bioreactor. 1081 16

A new engineering strain, Bacillus pumilus c172-14 (pBX 96), was obtained by introducing the pBX 96 plasmid, which carries the alpha-amylase amy gene, into the host strain of alkalophilic Bacillus pumilus c172 via transformation. The newly constructed strain was found to express the amy gene and could use starch instead of glucose or starch hydrolysate as carbon source for its fermentation of alkaline protease. The pBX 96 plasmid in the new host was found to be segregationally and structurally stable. The expression of amy gene did not affect the host strain's resistance to bacteriophages. Moreover, the level of alkaline protease was improved significantly compared with the parent strain. The constructed strain gave a maximum alkaline protease activity of 14,014 U/ml in shaking flask after 48 h cultivation when growing in a medium containing 6% corn meal, 4% soybean flour, 0.4% Na2HPO4, 0.03% KH2PO4, 0.02% MgCl2, 0.3% CaCl2, 0.25% Na2CO3, 0.1% glucose, and 20 microg/ml kanamycin (pH 7.0). The optimal pH value and temperature of the alkaline protease were 11.0 and 40 degrees C, respectively. This enzyme was stable over a pH range of 8-11. Its residual activity remained at 100% when treated under a temperature of less than 45 degrees C for 30 min. The corresponding residual activity reduced to 65% of its optimal value at 60 degrees C for 30 min. The alkaline protease was a kind of serine protease, which was demonstrated by the complete inactivation by PMSF (1 mM). This newly constructed strain will be useful in the alkaline protease industry.
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PMID:Fermentation of starch for enhanced alkaline protease production by constructing an alkalophilic Bacillus pumilus strain. 1169 13

A robust and reliable method was developed to measure resistant starch (RS), i.e., starch that enters the large intestine. In vivo conditions were reflected as much as possible while a user-friendly format was maintained. Parameters investigated included a-amylase concentration, pH of incubation, maltose inhibition of alpha-amylase, the need for amyloglucosidase inclusion, the effect of shaking and stirring on determined values, and problems in recovering and analyzing the RS-containing pellet. The RS values obtained were in good agreement with published in vivo data. An interlaboratory evaluation of the method has been completed (First Action Method 2002.02).
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PMID:Measurement of resistant starch. 1208 59

The optimization of cultural conditions for Beta-glucanase production by Bacillus subtilis ZJF-1A5 was investigated in flask trials. Temperature had great effect on Beta-glucanase production which maximized at optimal temperature of 37 degrees C and decreased significantly when temperature was over 37 degrees C. Charge quantity affected Beta-glucanase production significantly. Adding oxygen vector N-dodecane or acetic ether benefited Beta-glucanase production, but it depended on the concentration and charge quantity. The results of fractional factorial design showed that age and size of inoculum and shaking speed were the key factors affecting Beta-glucanase production and the cultivation time span to reach the highest Beta-glucanase activity. The optimal cultural conditions for Beta-glucanase production obtained with CCD were as follows: inoculum age and size (16 h, 3.82%(v/v)), shaking speed 210 r/min, charge quantity of 30 mL in 250 mL flask and initial pH 7.0, cultured at 37 degrees C for 50 h. Repeated experimental results accorded with those predicted by a second-order polynomial model. The amount of Beta-glucanase, Alpha-amylase and neutral protease produced by B subtilis ZJF-1A5 was associated partially with cell growth. Those three enzymes' activities increased following the cell growth and increased significantly when cells entered the stationary phase.
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PMID:Optimization of cultural conditions for thermostable beta-1,3-1,4-glucanase production by Bacillus subtilis ZJF-1A5. 1456 89

There is considerable interest in determining the added levels of the natural dye annatto in foods like snack products, particularly because they are mostly consumed by young people. The objective was to use response surface methodology to develop a new method to analyse annatto in extruded snacks. A pretreatment of the samples was necessary, digesting the ground sample with alpha-amylase at room temperature. The pigment was extracted by shaking with ethyl acetate at room temperature, eight extractions being necessary for completion extract the pigment. Lipids were removed by alkaline saponification. Under these conditions, 100% of the bixin was converted into norbixin, which was then quantified by high-performance liquid chromatography. The method had a mean recovery of 97% and a coefficient of variation for duplicate analysis of 1%. Using this method, of the 13 commercial samples analysed, a parmesan cheese-flavoured snack product showed the highest level of dye expressed as norbixin (15.5 mg kg(-1)), whilst other brands of onion-flavoured snack products had the lowest levels (0.7 and 0.4 mg kg(-1), respectively).
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PMID:Novel method for the determination of added annatto colour in extruded corn snack products. 1475 34

We cultivated a filamentous fungus, Aspergillus oryzae IAM 2706 by three different cultivation methods, i.e., shaking-flask culture (SFC), agar-plate culture (APC), and membrane-surface liquid culture (MSLC), to elucidate the differences of its behaviors by different cultivation methods under the same media, by measuring the growth, secretion of proteases and alpha-amylase, secreted protein level, and gene transcriptional profile by the DNA microarray analysis. The protease activities detected by MSLC and APC were much higher than that by SFC, using both modified Czapek-Dox (mCD) and dextrin-peptone-yeast extract (DPY) media. The alpha-amylase activity was detected in MSLC and APC in a much larger extent than that in SFC when DPY medium was used. On the basis of SDS-PAGE analyses and N-terminal amino acid sequences, 6 proteins were identified in the supernatants of the culture broths using DPY medium, among which oryzin (alkaline protease) and alpha-amylase were detected at a much higher extent for APC and MSLC than those for SFC while only oryzin was detected in mCD medium, in accordance with the activity measurements. A microarray analysis for the fungi cultivated by SFC, APC, and MSLC using mCD medium was carried out to elucidate the differences in the gene transcriptional profile by the cultivation methods. The gene transcriptional profile obtained for the MSLC sample showed a similar tendency to the APC sample while it was quite different from that for the SFC sample. Most of the genes specifically transcribed in the MSLC sample versus those in the SFC sample with a 10-fold up-regulation or higher were unknown or predicted proteins. However, transcription of oryzin gene was only slightly up-regulated in the MSLC sample and that of alpha-amylase gene, slightly down-regulated.
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PMID:Cultivation characteristics and gene expression profiles of Aspergillus oryzae by membrane-surface liquid culture, shaking-flask culture, and agar-plate culture. 2015 76

Conglomerates may form at the bottom of bottles containing not-from-concentrate (NFC) cloudy apple juice obtained from a single apple cultivar. Since dissociation or dispersion of these conglomerates cannot be achieved by shaking, the juice appears unsightly, and consumers can mistake the product as being spoilt. Juice from Elstar and Braeburn apples was treated with selected enzymes (gluco-amylase, alpha-amylase, pectin lyase, polygalacturonase and protease) and by using bentonite and a separator. The effects of these treatments on the formation of conglomerates in cloudy apple juice have been studied, and the treatment effects on turbidity and cloud stability have been documented. Conglomerates were observed in juices treated with protease or bentonite and in control juices. However, no conglomerates were observed in the juice samples treated with gluco-amylase, alpha-amylase, pectin lyase or pectin lyase in combination with polygalacturonase. The results obtained after treatment with a separator were not consistent. It is supposed that carbohydrate fractions play a more important role in conglomerate formation than proteins and phenols. Gluco-amylase and alpha-amylase treatment also provides good cloud stability, and thus can be a suitable method for preventing conglomerate formation in single-cultivar NFC cloudy apple juice.
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PMID:Prevention of conglomerate formation in not-from-concentrate single-cultivar cloudy apple juice by using different treatment methods. 2323 59