Temperature alternation by an on-chip microheater to reveal enzymatic activity of beta-galactosidase at high temperatures.

نویسندگان

  • Hideyuki F Arata
  • Yannick Rondelez
  • Hiroyuki Noji
  • Hiroyuki Fujita
چکیده

A method to measure enzymatic activity at high temperatures by rapid temperature alternation of a microreactor with a microheater is proposed. On-chip microreactor and microheater were integrated on a glass plate by MEMS technology; this microheater can control the temperature of the microreactor with a response speed of 34.2 and 31.5 K/s for temperature rise and fall, respectively, with an accuracy of 3 degrees C. The enzyme, beta-galactosidase, was revealed to survive short exposure (4-s pulses) to temperatures above that which would "normally" denature them. Its activity at 60 degrees C was revealed to be approximately 4 times greater than that at room temperature. This method not only gives new kinetic information in biochemistry but also enables application in highly sensitive biosensors.

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

β-galactosidase Production by Aspergillus niger ATCC 9142 Using Inexpensive Substrates in Solid-State Fermentation: Optimization by Orthogonal Arrays Design

Background: Enzymatic hydrolysis of lactose is one of the most important biotechnological processes in the food industry, which is accomplished by enzyme β-galactosidase (β-gal, β-D-galactoside galactohydrolase, EC 3.2.1.23), trivial called lactase. Orthogonal arrays design is an appropriate option for the optimization of biotechnological processes for the production of microbial...

متن کامل

Bioaffinity Based Immobilization of Almond (Amygdalus communis) b-galactosidase on Con A-layered Calcium Alginate-cellulose Beads: Its Application in Lactose Hydrolysis in Batch and Continuous Mode

In this study, immobilization of partially purified almond (Amygdalus communis) β-galactosidase on Con A layered calcium alginate-cellulose beads was investigated. Immobilized β-galactosidase retained 72% of theinitial activity after crosslinking by glutaraldehyde. Both soluble and immobilized enzyme exhibited the samepH and temperature optima at pH 5.5 and 50ºC, respectively. Howev...

متن کامل

PRODUCTION OF BETA-GALACTOSIDASE IN SUBMERGED MEDIA BY ASPERGILLUS ORYZAE, PTCC 5 163

Different compositions of liquid media were used for their efficacy in the production of beta-galactosidase by Aspergillus oryzae in submerged fermentation. Enzyme production was improved by switching from lactose-based media to soybean meal- and wheat bran-based media. Further improvement was made when the soybean meal medium was supplemented with sodium nitrate and magnesium sulfate. Res...

متن کامل

In Vitro Evaluation of Protective Effect of Rutin on Acrylamide-Induced Cellular Senescence in NIH3T3 Cells

Background: Aging is one of the important factors in the development of age-related diseases. Acrylamide can be produced during carbohydrate-rich foods prepared at high temperatures. Recently, studies showed that acrylamide can induce cellular senescence. On the other hand, Rutin as a natural flavonoid, has a potent antioxidant activity. Objective: This study aims to evaluate the ptotective ef...

متن کامل

Activation of plasma membrane H(+)-ATPase and expression of PMA1 and PMA2 genes in Saccharomyces cerevisiae cells grown at supraoptimal temperatures.

During exponential growth at temperatures of 30 to 39 degrees C, the specific activity of H(+)-ATPase in the plasma membrane of Saccharomyces cerevisiae (assayed at the standard temperature 30 degrees C) increased with increases in growth temperature. In addition, the optimal temperature for in vitro activity of this ATPase was 42 degrees C. Therefore, the maximum values of ATPase activity were...

متن کامل

ذخیره در منابع من


  با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

عنوان ژورنال:
  • Analytical chemistry

دوره 77 15  شماره 

صفحات  -

تاریخ انتشار 2005