Інститут біохімії ім. О.В Палладіна НАН України

Palladin Institute of Biochemistry of the NAS of Ukraine

BIOTECHNOLOGY VOL. 1, No.4, 2008

QUANTITATIVE ASSAY OF CHROMIUM(III) IN YEAST CULTURES USING CHROMAZUROL S AND SURFACTANTS FOR MONITORING CHROMATE REMEDIATION PROCESSES T. M. Honchar, H. P. Ksheminska, І. О. Patsay, О. М. Huta, М. V. Gonchar

"Biotechnology" journal Vol. 1, No. 4, 2008
Р. 85-94, Bibliography 28, English.
Universal Decimal classification 543.062 : 546.7 : 579.2

QUANTITATIVE ASSAY OF CHROMIUM(III) IN YEAST CULTURES USING CHROMAZUROL S AND SURFACTANTS FOR MONITORING CHROMATE REMEDIATION PROCESSES
T. M. Honchar, H. P. Ksheminska, І. О. Patsay, О. М. Huta, М. V. Gonchar

Because microorganisms are regarded as a potential means for bioremediation of chromium compounds and the yeasts are considered to be the source of chromium pharmaceutical biocomplexes, the development of the methods for chromium assay in real biological samples is of a significant biotechnological importance. A new sensitive method for the photometric assay of total chromium in the form of Cr(III) in microbial cultures after their mineralization was developed using the reaction with chromazurol S in the presence of the surfactants, sodium dodecyl sulfate (SDS) or cetyltrimethylammonium bromide (CTMAB).

The optimal values of chromazurol S and surfactants concentrations were determined to achieve the maximal sensitivity of the analysis. Molar extinction coefficients of the complexes in 0.2 M sulfuric acid were shown to equal 27 and 98.9 мМ–1•cm–1 in cases of using SDS and CTMAB, respectively. The latter variant of chromium (III) assay is 3 times more sensitive than diphenylcarbazide method. The threshold sensitivity of chromazurol method is approximately 100 and 25 ng of chromium per 1 ml of the final reaction mixture for two variants of the assay in cases of using SDS and CTMAB, respectively. A convenient and relatively fast method for mineralization of the biological samples was optimized using perhydrol and sulfuric acid. It was shown that the developed methods can be used for the monitoring of chromate-reduction processes in yeast cultures by the final product Cr(III), as well as for determination of the total chromium content in microbial cultures or in the cells after their mineralization.

Key words: chromium(ІІІ), chromazurol S, sodium dodecylsulfate, cetyltrimethylammonium bromide, photometric assay, yeast culture.

ENZYME CONDUCTOMETRIC BIOSENSOR FOR LACTOSE CONTENT DETERMINATION V. M. Pyeshkova, O. Y. Saiapina, O. O. Soldatkin, O. L. Kukla, S. V. Dzyadevych

"Biotechnology" journal Vol. 1, No. 4, 2008
Р. 76-84, Bibliography 20, Ukrainian.
Universal Decimal classification 577.15.543.555

ENZYME CONDUCTOMETRIC BIOSENSOR FOR LACTOSE CONTENT DETERMINATION
V. M. Pyeshkova, O. Y. Saiapina, O. O. Soldatkin, O. L. Kukla, S. V. Dzyadevych

Enzyme conductometric biosensor for lactose determination has been developed. Three-enzyme membrane (glucose oxidase, mutarotase, β-galactosidase) immobilized on the surface of conductometric transducer was used as biosensitive element. The analytical characteristic of the biosensor obtained were investigated. The time of lactose analysis in solution was 1–2 minutes, linear range was from 0,01 mM to 0,75 mM for glucose determination and from 0,01 mM to 1,25 mM for lactose determination. Dependence of responses of biosensor to substrate on pH, ionic strength, buffer capacity of work solution were studied, and the data of selectivity and storage stability were presented.

External view of conduktometric planar pectinate transducers
The developed conductometric biosensor is featured in high operational stability and signal reproducibility.
Key words: conductometric enzyme biosensor, lactose, glucose, glucose oxidase, mutarotase, β-galactosidase.

"Biotechnology" journal V. 1, No. 4, 2008
Р. 76-84, Bibliography 20, Ukrainian.
Universal Decimal classification 577.15.543.555

ENZYME CONDUCTOMETRIC BIOSENSOR FOR LACTOSE CONTENT DETERMINATION

V. M. Pyeshkova, O. Y. Saiapina, O. O. Soldatkin, O. L. Kukla, S. V. Dzyadevych

Enzyme conductometric biosensor for lactose determination has been developed. Three-enzyme membrane (glucose oxidase, mutarotase, β-galactosidase) immobilized on the surface of conductometric transducer was used as biosensitive element. The analytical characteristic of the biosensor obtained were investigated. The time of lactose analysis in solution was 1–2 minutes, linear range was from 0,01 mM to 0,75 mM for glucose determination and from 0,01 mM to 1,25 mM for lactose determination. Dependence of responses of biosensor to substrate on pH, ionic strength, buffer capacity of work solution were studied, and the data of selectivity and storage stability were presented.

BIOLUMINESCENT BACTERIA AS SENSOR ELEMENTS FOR THE HEAVY METALS IONS’ CONTENT DETECTION A. M. Zadorozhnyaya, T. G. Gruzina, S. M. Dibkova, Z. R. Ulberg

"Biotechnology" journal Vol. 1, No. 4, 2008
Р. 69-75, Bibliography 74, Ukrainian.
Universal Decimal classification 541.13:577.112.087

BIOLUMINESCENT BACTERIA AS SENSOR ELEMENTS FOR THE HEAVY
METALS IONS’ CONTENT DETECTION
A. M. Zadorozhnyaya, T. G. Gruzina, S. M. Dibkova, Z. R. Ulberg

It has been shown the doze-dependent inhibition influence of gold ions on Photobacterium phosphoreum В7071 (lux⁺) natural luminescence. These bacterial cells can serve as a sensor element for ionic gold content detection, for example, in wastewater of gold-mining and gold-processing manufactures for requirements compliance control. The Pseudomonas fragi T2(5) transconjugates, which contain hybrid plasmid (Zn^Rlux⁺) and which are capable of the high-specific of light emission as a result of the contact with zinc ions, have been derived by conjugate crossing method. It enables to use these transconjugates as sensor elements in the high-specific bioluminescent detection of qualitative and quantitative content of this metal in environmental objects.

Degree of inhibition of the cells natural luminescence of Photobacterium phosphoreum V7071 (lux⁺)

with ions of heavy metals:

1 —Au ions under concentration 0,2 mkg/ml (1,0 mkM);
2 —Au ions under concentration 0,5 mkg/ml (2,5 mkM);
3 —Zn ions under concentration 100 mkM;
4 —Со ions under concentration 100 mkM

Key words: bacteria, natural bioluminescence, induced bioluminescence, detection, gold, zinc, conjugate crossing.

Degree of inhibition of the cells natural
luminescence of
Photobacterium phosphoreum V7071 (lux⁺)

with ions of heavy metals:

1 —Au ions under concentration 0,2 mkg/ml (1,0 mkM);

2 —Au ions under concentration 0,5 mkg/ml (2,5 mkM);

3 —Zn ions under concentration 100 mkM;

4 —Со ions under concentration 100 mkM

MODIFICATION OF WHEAT STARCH BY DIFFERENT AMYLASES L. V. Kaprelyants, T. V. Shpirko, O. F. Pomazanova

"Biotechnology" journal Vol. 1, No. 4, 2008
Р. 64-68, Bibliography 19, Russian.
Universal Decimal classification 664.231: 577 15

MODIFICATION OF WHEAT STARCH BY DIFFERENT AMYLASES
L. V. Kaprelyants, T. V. Shpirko, O. F. Pomazanova

Effect of different preparations of α-amylase on the wheat starch and its fractions has been studied. Oligosaccharides with different molecular mass have been detected in the samples. It has been shown that hydrolysates with low glucose equivalent have the thermally reversible properties under the determined interrelation between low- and high-polymeric fractions of the starch.

Key words: starch, amylases, hydrolysate, enzymatic hydrolysis, oligosaccharides.

MICROCLONAL PROPAGATION OF UNGERNIA VICTORIS VVED. EX ARTJUSCHENKO V. А. Kunakh, L. P. Mozhylevska, O. M. Bublyk, I. V. Kolonina, V. I. Muzyka

"Biotechnology" journal Vol. 1, No. 4, 2008
Р. 57-63, Bibliography 13, Ukrainian.
Universal Decimal classification 615.222:581.143.6

MICROCLONAL PROPAGATION OF UNGERNIA VICTORIS VVED. EX ARTJUSCHENKO
V. А. Kunakh, L. P. Mozhylevska, O. M. Bublyk, I. V. Kolonina, V. I. Muzyka

Approaches to microclonal propagation of the rare medicinal plant Ungernia victoris both through the direct regeneration from the scale of bulb and via induction of regeneration from the long_term grown in vitro callus tissues have been developed for the first time ever. Conditions for multiplication and maintenance of in vitro generated microbulb-regenerants were specified as well.

Microclonal propagation of the plant U. victoris:

a - general view of one of the initial bulbs with mass of 120 g and age of about 45 years, reared in natural conditions (the southern slopes of Hissar Mountains, the Pamirs, Tajikistan);
б - primary kalyus at the 60-th day after isolation of a fragment of bulb scin;
в –mikrobulbs regeneration occuring in the primary eksplante together with kalyus creation;
г – organogenic passivated tissue culture spontaneously forming rootlike and teratomolike structures;
д - regenerants micropropagation: transplanted mikrobulb (dark color) in 5S3I medium forms new mikrobulbs (light color).

Key words: Ungernia victoris, microclonal propagation, direct plant regeneration, induction of plant regeneration.
© Palladin Institute of Biochemistry of National Academy of Sciences of Ukraine, 2008