scholarly journals Biochemical studies on the activity of δ-(l-α-aminoadipyl)-l-cysteinyl-d-valine synthetase from Streptomyces clavuligerus

1992 ◽  
Vol 283 (3) ◽  
pp. 691-698 ◽  
Author(s):  
J Zhang ◽  
S Wolfe ◽  
A L Demain

The enzyme activity of purified delta-(L-alpha-aminoadipyl)-L-cysteinyl-D-valine (ACV) synthetase from Streptomyces clavuligerus was studied biochemically. The dependence of ACV synthetase activity on reaction parameters, including substrates, cofactors, temperature and pH, were determined, resulting in a substantially increased enzyme activity. The activity is very labile to high temperature and is also unstable at acidic pH. The enzyme specificity is strict towards L-alpha-aminoadipate, but rather loose with respect to L-valine; certain modifications of L-cysteine can also be tolerated. Some unnatural tripeptides synthesized by ACV synthetase can be converted into bioactive compounds by isopenicillin N synthase. The only nutrient found to negatively affect ACV synthetase activity is phosphate, but various compounds such as thiol-blocking reagents and ATP-utilization products (AMP and pyrophosphate) are inhibitory to the enzyme.

1984 ◽  
Vol 222 (3) ◽  
pp. 789-795 ◽  
Author(s):  
C P Pang ◽  
B Chakravarti ◽  
R M Adlington ◽  
H H Ting ◽  
R L White ◽  
...  

Isopenicillin N synthetase was extracted from Cephalosporium acremonium and purified about 200-fold. The product showed one major protein band, coinciding with synthetase activity, when subjected to electrophoresis in polyacrylamide gel. An isopenicillin N synthetase from Penicillium chrysogenum was purified about 70-fold by similar procedures. The two enzymes resemble each other closely in their Mr, in their mobility on electrophoresis in polyacrylamide gel and in their requirement for Fe2+ and ascorbate for maximum activity. Preliminary experiments have shown that a similar isopenicillin N synthetase can be extracted from Streptomyces clavuligerus.


1984 ◽  
Vol 47 (2) ◽  
pp. 105-107 ◽  
Author(s):  
BARBARA P. KEOGH ◽  
G. PETTINGILL

An investigation was undertaken into the relationship between the enzyme activity of cells harvested from raw milk and time taken for age gelation (TAG) to occur in the milk after ultra-high-temperature processing. It was shown that there was no relationship between the TAG and the bacterial counts on milk agar at 30°C or 7°C nor was there a relationship between the counts and the level of enzyme activity of the harvested cells. There was, however, a significant correlation between the level of enzyme activity of the harvested cells and the TAG. When extra bovine leucocytes were added to raw milk before processing, the TAG was increased. This suggested that there was an inhibitory action of leucocytes in development of age gelation.


Author(s):  
Gaļina Zvaigzne ◽  
Daina Kārkliņa ◽  
Joerg-Thomas Moersel ◽  
Sasha Kuehn ◽  
Inta Krasnova ◽  
...  

Abstract Orange juices are an important source of bioactive compounds. Because of its unique combination of sensory attributes and nutritional value, orange juice is the world’s most popular fruit juice. Orange (Citrus sinensis) juice of Greek Navel variety was used in this study. The impact of Conventional Thermal Pasteurisation (94 °C/30') (CTP) and alternative Ultra-High Temperature (UHT) (130 °C/2') processing on bioactive compounds and antioxidant capacity changes of fresh Navel orange juice was investigated. Sensory attributes of processed juices were evaluated. Results showed that using technologies CTP and UHT orange juice Navel significantly changed vitamin C concentration in comparison with fresh orange juice. The highest concentration of antioxidants (vitamin C, total phenols, hesperidin and carotenoids) was observed in orange juice Navel produced by UHT technology. Sensory results indicated that characteristics of the orange juice obtained using UHT technology were more liked than the CTP heat treated juice. UHT technology emerges as an advantageous alternative process to preserve bioactive compounds in orange juice.


Parasitology ◽  
1973 ◽  
Vol 67 (2) ◽  
pp. 197-204 ◽  
Author(s):  
Madan M. Goil

Biochemical studies on the non-specific phosphomonoesterases have demonstrated the presence of acid phosphomonoesterase with maximum activity at pH 4·0 in Gastrodiscus aegyptiacus (enzyme I) and at pH 4·5 in the case of Fasdolopsis buski (enzyme II). The Km for ρ-nitrophenyl phosphate hydrolysis was 0·66 mM for enzyme I and 1·1 mM for enzyme II. Different concentrations of fluoride, arsenate, tartrate, tartaric acid, cysteine and copper brought about inhibition of both enzymes and magnesium, iodoaeetate, iodoacetamide and EDTA had no influence on either enzyme activity. Cobalt activated both enzymes while zinc inhibited enzyme I and strongly stimulated enzyme II.


It was shown recently by Zeile and Hellström (1930) that a strong preparation of catalase obtained from horse liver has the distinct absorption spectrum of a haematin compound. The concentration of this haematin compound in various fractions of their preparations, estimated as pyridine haemochromogen, was found to be proportional to the enzyme activity of these fractions. They have also shown that the factors which abolish or inhibit the activity of the enzyme, such as high temperature, alkali, acids, or the addition of small amounts of KCN or H 2 S, modify also the absorption spectrum of the haematin.


Blood ◽  
1985 ◽  
Vol 66 (5) ◽  
pp. 1068-1071 ◽  
Author(s):  
MA Baker ◽  
RN Taub ◽  
A Kanani ◽  
I Brockhausen ◽  
A Hindenburg

Granulocytes from patients with chronic myelogenous leukemia (CML) are morphologically identical to their normal counterparts but show marked differences in circulation patterns and in some membrane properties. We have previously shown that there is abnormal lectin binding to CML granulocytes, and aberrant sialylation of membrane glycoproteins. To examine the changes in sialylation of CML granulocytes further, we have studied membrane preparations from CML and normal granulocytes for specific sialyltransferase activity. Because sialyltransferase enzymes are specific for the configuration of the acceptor group, enzyme activity was assayed by measuring transfer of sialic acid from CMP-14C- sialic acid to substrates of defined structure. As compared with those of normal counterparts, CML extracts catalyzed a 50% higher overall rate of sialylation of asialofetuin, a substrate possessing both N- and O-linked acceptors. Studies of enzyme specificity utilizing porcine and ovine submaxillary mucins, antifreeze glycoprotein and alpha-1 acid glycoprotein as acceptors showed that the increased sialylation by CML extracts was due primarily to substrates with the O-linked Gal beta 1--- -3GaINAc acceptor group. These data suggest that sialyltransferase activity is increased in CML granulocytes compared to normal granulocytes and that the increased enzyme activity is specific for O- linked Gal beta 1----3GaINAc. This enzyme activity may be directly responsible for the abnormal membrane sialylation and pathophysiological behavior of these cells.


Author(s):  
Claudia Capusoni ◽  
Immacolata Serra ◽  
Silvia Donzella ◽  
Concetta Compagno

Phytic acid is an anti-nutritional compound able to chelate proteins and ions. For this reason, the food industry is looking for a convenient method which allows its degradation. Phytases are a class of enzymes that catalyze the degradation of phytic acid and are used as additives in feed-related industrial processes. Due to their industrial importance, our goal was to identify new activities that exhibit best performances in terms of tolerance to high temperature and acidic pH. As a result of an initial screening on 21 yeast species, we focused our attention on phytases found in Cyberlindnera jadinii, Kluyveromyces marxianus, and Torulaspora delbrueckeii. In particular, C. jadinii showed the highest secreted and cell-bound activity, with optimum of temperature and pH at 50°C and 4.5, respectively. These characteristics suggest that this enzyme could be successfully used for feed as well as for food-related industrial applications.


Author(s):  
Ibrahim M. Abu-Reidah ◽  
Amber Critch ◽  
Charles Manful ◽  
Amanda Rajakaruna ◽  
Natalia Prieto Vidal ◽  
...  

Mushrooms have long rich history in folk medicine, traditional and functional foods due to high content of dietary myco-nutrients. Currently, there is increased interest in finding appropriate food-grade green ex-traction systems capable of extracting these bioactive compounds from dietary mushrooms for applica-tions in various food, pharmacological or nutraceutical formulations. Herein, we evaluated a modified Swiss water process (SWP) method using alkaline and acidic pH at low and high temperature under pressurized conditions as a suitable green food grade solvent to obtained extracts enriched with my-co-nutrients (dietary phenolics, total antioxidants (TAA), vitamins, and minerals) from Chaga. Ultra-high performance liquid chromatography coupled to high resolution accurate mass tandem mass spectrometry (UHPLC-HRAMS-MS/MS) was used to assess the phenolic compounds and vitamin levels in the extracts, while inductively coupled plasma mass spectrometry (ICP-MS) was used to determine the mineral con-tents. Over twenty phenolic compounds were quantitatively evaluated in the extracts and the highest total phenolic content and antioxidant activity was observed at pH11.5 at 100°C. The most abundant phenolic compounds present in Chaga extracts included phenolic acids such as protocatechuic acid 4-glucoside (0.7-1.08µg/mL), syringic acid (0.62-1.18µg/mL), and myricetin (0.68-1.3µg/mL). Vitamins are being reported for the first time in Chaga. pH 2.5 at 100°C treatment shows superior effects in extracting the B vitamins whereas pH 2.5 at 60 and 100°C treatments were outstanding for extraction of total fat-soluble vitamins. Vitamin E content was the highest for the fat-soluble vitamins in the Chaga extract under acidic pH (2.5) and high temp. (100°C) and ranges between 50 to 175 µg/100g Chaga. Antioxidant minerals ranged from 85.94 µg/g (pH7 at 100°C) to 113.86 µg/g DW (pH2.5 at 100°C). High temperature 100°C and a pH of 2.5 or 9.5. The treatment of pH11.5 at 100°C was the most useful for recovering phenolics and antioxidants from Chaga including several phenolic compounds reported for the first time in Chaga. SWP is being proposed herein for the first time as a novel, green food-grade solvent system for the extraction of myco-nutrients from Chaga and have potential applications as a suitable approach to extract nutrients from other matrices. Chaga extracts enriched with bioactive myconutrients and antioxidants may be suitable for further use or applications in the food and nutraceutical industries.


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