NANOTECHNOLOGY IN ENZYME TECHNOLOGY

• PRESENTED TO :-

Dr PARVEEN PAHUJA • PRESENTED BY :-

SUDHA CHIB

130181107

M.Sc (H) BT 2

DEFINATION• Nanotechnology is the study

of manipulating matter on anatomic scale.

• Nanotechnology refers to theconstructing and engineeringof the functional systems atvery micro level or we can sayat atomic level.

• A Nanometer is one billionthof a meter, roughly the widthof three or four atoms. Theaverage human hair is about25,000 nanometers wide.

HISTORY

• The first ever concept was presentedin 1959 by the famous professor ofphysics Dr. Richard P.Feynman.

• Invention of the scanning tunnelingmicroscope in 1981 and the discoveryof fullerene(C60) in 1985 lead to theemergence

of nanotechnology.

• The term “Nano-technology" hadbeen coined by Norio Taniguchi in1974

SELECTED NANOPARTICLES

• Carbon, ≥99% trace metals basis

• Diamond, ≥97% trace metals basis

• Diamond, ≥95% trace metals basis

• Titanium(IV) oxide, mixture of rutile and anatase, 99.9% trace metals basis

• Aluminum oxide

• Cerium(IV) oxide, >99.95% trace metals basis

ENZYME IMMOBILISATION

• Application of nanomaterials as novel supporting materials for enzyme immobilisation has generated incredible interest in the biotechnology community.

• These robust nanostructured forms, such as nanoparticles, nanofibres, nanotubes, nanoporous, nanosheets, and nanocomposites, possess a high surface area to volume ratios that can cause a high enzyme loading and facilitate reaction kinetics, thus improving biocatalytic efficiency for industrial applications.

• The current status of versatile nanomaterial support for biofuel production employing cellulases and lipases is described in details

• nanomaterials will become an integral part of sustainable bioenergy production.

NANOBIOSENSORS

• A biosensor is defined as a device that uses specific biochemical reactions mediated by isolated enymes, immunosystems etc .to detect chemical compound

• Nanosensors with immobilized bioreceptorprobes that are selective for target analytemolecules are called nanobiosensors

APPLICATIONS

• Detection of microorganism in various sample

• Monitoring of metabolities in body fluids and detection of tissue pathology such as cancer

SINGLE ENZYME NANOPARTICLES

• SEN (single enzyme nanoparticles )

• Enzyme lead short and brutal lives ,to increase the enzymes longevity and versatility, a a team at department of Energy’s Pacific Northwest , National Laboratory in Richlad caged single enzyme to create a new class of catalysts called SENs

• The nanostructure protects the catalyst, allowing it to remain active for several months

• Kim and Grate , working in te W.R Wiley Environmental molecular sciences laboratory modified a common protein splitting enzyme called alpha chymotrypsin

ENHANCEMENT OF ENZYME ACTIVITY AND THERMOSTABILITY

Study on Impaired Pectate Lyase from AttenuatedMacrophominaphaseolina in Presence of Hydroxyapatite Nanoparticle

Hydroxyapatite nanoparticles (NP) can not only act as a chaperon (by imparting thermostability) but can serve as a synthetic enhancer of activity of an isolated extracellular pectate lyase (APL) with low native state activity.

The purified enzyme showed feeble activity at 50°C and pH 5.6. However, on addition of 10.5 µg/ml of hydroxyapatite nanoparticles(NP), APL activity increased 27.7 fold with a 51 fold increase in half-life at a temperature of 90°C as compared to untreated APL.

The upper critical temperature for such compensation was elevated from 50°C to 90°C in presence of NP.

EnzMet (Enzyme Metallography)

• EnzMet (Enzyme Metallography) is a new biological labeling and staining method developed at Nanoprobes.

• It uses a targeted enzymatic probe with a novel metallographic substrate to provide a quantum leap in staining clarity over conventional chromogenic and fluorescent substrates.

• EnzMet™ has proven highly sensitive both for in situ hybridization (ISH), where it readily visualizes endogenous copies of single genes, and immunohistochemistry (IHC) detection.

APPLICATIONS

In situ hybridization (ISH)

Immunohistochemistry (IHC)

Electron Microscopy

Light and Electron Correlative

Microscopy

Nanowires

HER2 staining in HER2-amplified tissue from a human breast cancer biopsy. Dr.

Raymond R. Tubbs, Cleveland Clinic.

NANOTECHNOLOGY TO HARNESS THE NATURAL LIGHT PRODUCED BY

FIREFLIES• By designing a way to chemically attach genetically

manipulated luciferase enzymes directly to the surface of nanorods, scientists at Syracuse University found a new way to harness the natural light produced by fireflies.

• Fireflies produce light through a chemical reaction between luciferin and it’s counterpart, the enzyme luciferase.

• In Maye’s laboratory, the enzyme is attached to the nanorod’s surface; luciferin, which is added later, serves as the fuel.

• The energy that is released when the fuel and the enzyme interact is transferred to the nanorods, causing them to glow. The process is called Bioluminescence Resonance Energy Transfer (BRET).

• The nanorods are composed of an outer shell of cadmium sulfide and an inner core of cadmium seleneide.

• http://www.azonano.com/article.aspx?ArticleID=736

• http://omicsonline.org/2153-0777/2153-0777-2-e114.pdf

• http://www.nanoprobes.com/products/EnzMet-SISH-enzyme-metallography-for-ISH-and-IHC.html

• http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0063567

• http://www.che.udel.edu/research_groups/wilfred/Current%2013%20final.pdf