This document discusses the use of nanotechnology in enzyme technology. It defines nanotechnology as manipulating matter at the atomic scale. Some key points discussed include immobilizing enzymes onto nanomaterials like nanoparticles to improve biocatalytic efficiency for applications. Nanoparticles are also shown to enhance enzyme activity and thermostability. Single enzyme nanoparticles are created by caging individual enzymes within nanostructures to increase their longevity. New techniques like EnzMet use enzymes with metallographic substrates to provide high clarity staining for applications in immunohistochemistry and electron microscopy. Harnessing firefly bioluminescence using luciferase enzymes attached to nanorods is also summarized.

1. NANOTECHNOLOGY IN ENZYME
TECHNOLOGY
• PRESENTED TO :-
Dr PARVEEN PAHUJA • PRESENTED BY :-
SUDHA CHIB
130181107
M.Sc (H) BT 2

2. DEFINATION
• Nanotechnology is the study
of manipulating matter on an
atomic scale.
• Nanotechnology refers to the
constructing and engineering
of the functional systems at
very micro level or we can say
at atomic level.
• A Nanometer is one billionth
of a meter, roughly the width
of three or four atoms. The
average human hair is about
25,000 nanometers wide.

3. HISTORY
• The first ever concept was presented
in 1959 by the famous professor of
physics Dr. Richard P.Feynman.
• Invention of the scanning tunneling
microscope in 1981 and the discovery
of fullerene(C60) in 1985 lead to the
emergence
of nanotechnology.
• The term “Nano-technology" had
been coined by Norio Taniguchi in
1974

5. 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

6. 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.

8. 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 bioreceptor
probes that are selective for target analyte
molecules are called nanobiosensors

9. APPLICATIONS
• Detection of microorganism in various sample
• Monitoring of metabolities in body fluids and
detection of tissue pathology such as cancer

11. 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

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

13. ENHANCEMENT OF ENZYME ACTIVITY
AND THERMOSTABILITY
Study on Impaired Pectate Lyase from AttenuatedMacrophomina
phaseolina 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.

14. 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.

15. APPLICATIONS
In situ hybridization (ISH)
Immunohistochemistry (IHC)
Electron Microscopy
Light and Electron Correlative
 Microscopy
Nanowires

16. HER2 staining in HER2-amplified tissue
from a human breast cancer biopsy. Dr.
Raymond R. Tubbs, Cleveland Clinic.

17. 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.

18. • 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.

19. • 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%2F1
0.1371%2Fjournal.pone.0063567
• http://www.che.udel.edu/research_groups/wilfre
d/Current%2013%20final.pdf