A Comparative Biochemical Characterization of Microbial Transglutaminases: Commercial vs. a Newly Isolated Enzyme from <Emphasis Type="Italic">Streptomyces</Emphasis> Sp.

A new microbial transglutaminase (MTGase or MTG, EC 2.3.2.13) from a Streptomyces sp. strain isolated from Brazilian soil samples was characterized in crude and purified forms. The aim of this work is to provide relevant information about a new transglutaminase and to compare its characteristics with the well-known commercial transglutaminase from Ajinomoto Co. Inc. (Activa® TG-BP). The enzyme from Streptomyces sp., in both crude and pure forms, exhibited optimal activity in the 6.0–6.5 pH range and at 35–40°C. The results for the commercial enzyme were the same. A second maximum of activity was observed at pH 10.0 with both the crude Streptomyces sp. enzyme and the commercial enzyme. This interesting fact has not been reported in the literature previously. The fact that this second maximum of activity does not appear on the purified form of the enzyme may suggest the presence of an isoenzyme on the crude extract. All of the enzymes tested were stable over the pH range from 4.5 to 8.0 and up to 45°C. The decline in activity of the commercial transglutaminase above 45°C and pH 8.0 was more gradual. The activities of all the MTG samples were independent of Ca⁺² concentration, but they were elevated in the presence of K⁺, Ba²⁺, and Co²⁺ and inhibited by Cu²⁺ and Hg²⁺, which suggests the presence of a thiol group in the MTG’s active site. The purified enzyme presented a K _m of 6.37 mM and a V _max of 1.7 U/mL, while the crude enzyme demonstrated a K _m of 6.52 mM and a V _max of 1.35 U/mL.     

Evaluation of Norrish’s Equation for Correlating the Water Activity of Highly Concentrated Solutions of Sugars,Polyols, and Polyethylene Glycols

Norrish’s equation, a_\textw = X_\textw exp( - KX²_\texts )a_{{\text{w}}} = X_{{\text{w}}} \exp {\left( { - KX^{2}_{{\text{s}}} } \right)}, where a _w is water activity, X _w and X _s are molar fractions of water and solute, respectively, and K is the correlating constant, has been widely used to predict a _w of aqueous nonelectrolyte solutions in connection with development of intermediate moisture foods, i.e., food having a _w ≥ 0.85. Present work evaluated the ability of Norrish’s equation to model the water activity of solutions of sugars, polyols, and some polyethylene glycols, in a wide range of concentration, i.e., from low to highly concentrated solutions. For sugar and polyols, a relatively small modification of the “most accepted” literature parameters K allowed the fitting of the data for the wide range of solute concentrations corresponding to a range of a _w from 0.99 to about 0.3 for same solutes. However, a modified Norrish’s model needs to be used to model the behavior of polyethylene glycols 400 and 600 up to water activities as low as 0.5.     

Extracts of Mexican Oregano (<Emphasis Type="Italic">Lippia berlandieri</Emphasis> Schauer) with Antioxidant and Antimicrobial Activity

Antimicrobial activity of fractions obtained from Mexican oregano (Lippia berlandieri Schauer) chloroform extract was tested by growth inhibition against Escherichia coli, Staphylococcus aureus and Bacillus cereus, and antioxidant capacity was tested by inhibition of linoleic acid oxidation. Fractions were obtained by differences in polarity or structure (phenolic and non-phenolic fraction). Gram-positive organisms were more susceptible to Mexican oregano extracts. Fraction 3 (by polarity) and phenolic fractions I, II, III, IV and V were the extracts with higher antimicrobial activity. The non-phenolic fraction had effect against B. cereus. Polarity fraction 5 and phenolic Fraction II had a high antioxidant capacity; a 0.08% concentration of fraction 5 had a similar effect as butylated hydroxytoluene at 0.01% concentration. Fractions of Mexican oregano with different polarity and functional groups had antioxidant and antimicrobial activity and can be used in a variety of applications.     

Alternative woods for aging distillates-an insight into their phenolic profiles and antioxidant activities

Work has commenced on the study and characterization of wood native species (oak, chestnut, cherry, alder, ash, and beech), employed to a minor extent in enology, for their application in the elaboration of distillates. To this end, furanic and low molecular weight phenol compounds, total phenolic content, and antioxidant activity in alcoholic extracts by maceration of wood chips at 2 toasting levels were studied. Significant differences between samples, technologically relevant, were due both to species and toasting time. Furthermore, extracts showed interesting antioxidant activities equivalent in some cases to than those reported for alcoholic beverages aged in wood.     

Cation/H+ antiporters mediate potassium and sodium fluxes in Pichia sorbitophila. Cloning of the PsNHA1 and PsNHA2 genes and expression in Saccharomyces cerevisiae

Pichia sorbitophila grows rapidly in the presence of very high NaCl concentrations. Under these conditions, even when the K(+) concentration is low, P. sorbitophila cells can maintain low Na(+) and high K(+) contents. This remarkable capacity of P. sorbitophila fails when the external pH is not acidic. This indicates that Na(+) efflux is mediated by an electroneutral Na(+)/H(+) antiporter. We have cloned and sequenced two genes designated as PsNHA1 and PsNHA2, which probably encode two antiporters of this type. The genes present high similarity with the corresponding genes from other yeasts. The heterologous expression of PsNHA1 or PsNHA2 in a Saccharomyces cerevisiae mutant lacking the Na(+) efflux systems and sensitive to high concentrations of Na(+) and K(+) rescued the tolerance and the ability to extrude both cations. The Accession Nos of the sequenced DNA fragments are: PsNHA1, AJ496431; PsNHA2, AJ496432. (TC 2.A.36) Copyright 2002 John Wiley & Sons, Ltd.     

Influence of starter and nonstarter on the formation of biogenic amine in goat cheese during ripening

Two commercial starters were investigated for their potential ability to decarboxylate amino acids during goat cheese ripening. Two batches of goat cheese were produced with identical pasteurized milk but different starter cultures. One of them contained Lactococcus lactis subsp. lactis and Lactococcus lactis subsp. cremoris and the other Lactococcus lactis subsp. lactis. The amine contents, microbial counts, proteolysis-related parameters, pH, total solids and salt content were studied in raw materials and cheeses. In raw materials, polyamines were the prevailing amines, whereas the main amines in cheeses were putrescine, tryptamine and, in particular, tyramine (94.59 mg/kg). Aerobic mesophilic microorganisms and Lactococcus counts increased throughout ripening, while Enterobacteriaceae were no longer detectable in cheese after 30 days of ripening. Amine concentration rose during cheese ripening in both batches. Moreover, the decarboxylase activity of microorganisms isolated from samples during cheese ripening was assayed and discussed.     

Nanofiltration for the recovery of caustic cleaning-in-place solutions: robustness towards large variations of composition

In the dairy industry re-use and multi-use cleaning-in-place (CIP) systems are operated by circulating chemicals and water without taking the equipment apart. The solutions, which become polluted due to the removal of fouling compounds, are drained periodically when they are considered to be too polluted. This work shows the large variations in composition (pollution, surface tension, etc) of the industrial caustic solutions coming from milk standardization and pasteurization plant CIP throughout their life time (7 days) and from 1 week to another. The work is also intended to show how nanofiltration (1 kg mol(-1) molecular weight cut-off) was robust and performed well, with good recovery of caustic solutions, even when faced with large variations of solutions composition: high caustic yield, permeation flux (J) in the range 42-110 l h(-1) m(-2), average chemical oxygen demand (COD) reduction equal to 0.58 and low surface tension change. Equations have been established for the prediction of J as a function of initial membrane hydraulic resistance (Rm) caustic concentration, volume reduction ratio (VRR) and initial soluble COD. When VRR increased, both J and pollution retention decreased despite the increase in irreversible fouling induced by the increase of soluble pollution concentration in retentate. The higher the initial soluble COD, the sharper the decrease in J vs. VRR. Since irreversible fouling was usually small (0.1-3.4 x 10(13) m(-1), that is to say of the same order of magnitude as Rm), the membrane cleaning could be efficiently performed by using single phase sodium hypochlorite alternately with a more expensive acid-base cleaning sequence. The obtained permeate was a clear regenerated cleaning solution with low soluble COD (0.2-3.5 g/l) and surface tension (56-30 mJ m(-2)) which could be successfully exploited owing to its cleaning potential.     

Nudging hair shedding by antidandruff shampoos. A comparison of 1% ketoconazole, 1% piroctone olamine and 1% zinc pyrithione formulations

Hair shedding and hair thinning have been reported to be affected by dandruff and seborrhoeic dermatitis. The present study was conducted in 150 men presenting with telogen effluvium related to androgenic alopecia associated with dandruff. They were randomly allocated to three groups receiving each one of the three shampoos in the market containing either 1% ketoconazole (KTZ), 1% piroctone olamine (PTO) or 1% zinc pyrithione (ZPT). Shampoos had to be used 2-3 times a week for 6 months. Hair shedding during shampoo was evaluated semiquantitatively. Hair density on the vertex was evaluated on photographs using a Dermaphot. Trichograms were used for determining the anagen hair percentage and the mean proximal hair shaft diameter using computerized image analysis. The sebum excretion rate (SER, mug cm(-2) h(-1)) was also measured using a Sebumeter. The three treatments cleared pruritus and dandruff rapidly. At end point, hair density was unchanged, although hair shedding was decreased (KTZ: -17.3%, PTO: -16.5%, ZPT: -10.1%) and the anagen hair percentage was increased (KTZ: 4.9%, PTO: 7.9%, ZPT: 6.8%). The effect on the mean hair shaft diameter was contrasted between the three groups of volunteers (KTZ: 5.4%, PTO: 7.7%, ZPT: -2.2%). In conclusion, telogen effluvium was controlled by KTZ, PTO and ZPT shampoos at 1% concentration. In addition, KTZ and PTO increased the mean hair shaft thickness while discretely decreasing the sebum output at the skin surface.