Team:IIT Madras/Dry lab/Circuitry

From 2011.igem.org

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<font face="calibri" color="#080000"> One of the challenging task was to establish the lighting setup which would power the proteorhodopsin  in presence of retinal to carry its H+ pumping activity in the carbon deficient condition. On rigorous searching  we came across a research paper (reference and relevant extract mentioned below) which clearly defined the wavelenght and intensity to maintain the proton motive force (pmf).<br/>
<font face="calibri" color="#080000"> One of the challenging task was to establish the lighting setup which would power the proteorhodopsin  in presence of retinal to carry its H+ pumping activity in the carbon deficient condition. On rigorous searching  we came across a research paper (reference and relevant extract mentioned below) which clearly defined the wavelenght and intensity to maintain the proton motive force (pmf).<br/>
<b>Refernce : Light-powering Escherichia coli with proteorhodopsin  Jessica M. Walter, Derek Greenfield, Carlos Bustamante, and Jan Liphardt . Contributed by Carlos Bustamante, December 13, 2006 </b><br/><br/>
<b>Refernce : Light-powering Escherichia coli with proteorhodopsin  Jessica M. Walter, Derek Greenfield, Carlos Bustamante, and Jan Liphardt . Contributed by Carlos Bustamante, December 13, 2006 </b><br/><br/>
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Prior to this paper the role of light in powering cells containing proteorhodopsin and participation in ocean energy fluxes remained largely unclear. This paper makes an attempt to show that when cellular respiration is inhibited by depleting oxygen or by the respiratory poison azide, Escherichia coli cells expressing PR become light-powered.<br/>
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Prior to this paper the role of light in powering cells containing proteorhodopsin and participation in ocean energy fluxes remained largely unclear. This paper makes an attempt to show that when cellular respiration is inhibited by depleting oxygen or by the respiratory poison azide, Escherichia coli cells expressing PR become light-powered.
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<br/><br/>
<h3><font face="arial" color="#980000 "><b><u>Instrumentation</u></b></font></h3>
<h3><font face="arial" color="#980000 "><b><u>Instrumentation</u></b></font></h3>
Power density values for <font color="#00CC33"><b>green light</b></font> refer to the power density passed by a D540/25ϫ filter originating at a 175 W Xenon bulb .The sample chamber  was periodically illuminated with bright green light (160mW/cm2) coinciding with the maximum of PR’s absorption spectrum, 525 nm. At 525 nm the sample was observed to show maximum absorption (fig.1).<br/>
Power density values for <font color="#00CC33"><b>green light</b></font> refer to the power density passed by a D540/25ϫ filter originating at a 175 W Xenon bulb .The sample chamber  was periodically illuminated with bright green light (160mW/cm2) coinciding with the maximum of PR’s absorption spectrum, 525 nm. At 525 nm the sample was observed to show maximum absorption (fig.1).<br/>
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Based on the above fact we decided to use green LED of dominant wavelenght of 525 nm.<br/>
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Based on the above fact from the above given reference we decided to use green LED of dominant wavelenght of 525 nm.
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<br/><br/>
<h3><font face="arial" color="#980000 "><b><u>Diagrams</u></b></font></h3>
<h3><font face="arial" color="#980000 "><b><u>Diagrams</u></b></font></h3>
<img src="https://static.igem.org/mediawiki/2011/8/87/H11.png" width="340" height="250"/>
<img src="https://static.igem.org/mediawiki/2011/8/87/H11.png" width="340" height="250"/>
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<img src="https://static.igem.org/mediawiki/2011/9/9f/H13.png" width="340" height="250"/>
<img src="https://static.igem.org/mediawiki/2011/9/9f/H13.png" width="340" height="250"/>
<img src="https://static.igem.org/mediawiki/2011/c/cf/H14.png" width="340" height="250"/><br/>
<img src="https://static.igem.org/mediawiki/2011/c/cf/H14.png" width="340" height="250"/><br/>
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<font size=2><u><b>fig(1)</b> & <b>fig(2)</b> Copyright@Light-powering Escherichia coli with proteorhodopsin  Jessica M. Walter, Derek Greenfield, Carlos Bustamante, and Jan Liphardt. Contributed by Carlos Bustamante, December 13, 2006 </u></font><br/><br/>
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<font size=2><u><b>fig(1)</b> & <b>fig(2)</b> Copyright@Light-powering Escherichia coli with proteorhodopsin  Jessica M. Walter, Derek Greenfield, Carlos Bustamante, and Jan Liphardt. Contributed by Carlos Bustamante, December 13, 2006 </u></font>
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<br/><br/>
<h3><font face="arial" color="#980000 "><b><u>Specifications</u></b></font></h3>
<h3><font face="arial" color="#980000 "><b><u>Specifications</u></b></font></h3>
<ol>
<ol>

Revision as of 11:47, 5 October 2011

bar iGEM 2011 - Home Page Indian Institute of Technology - Madras











GREEN LANTERN

One of the challenging task was to establish the lighting setup which would power the proteorhodopsin in presence of retinal to carry its H+ pumping activity in the carbon deficient condition. On rigorous searching we came across a research paper (reference and relevant extract mentioned below) which clearly defined the wavelenght and intensity to maintain the proton motive force (pmf).
Refernce : Light-powering Escherichia coli with proteorhodopsin Jessica M. Walter, Derek Greenfield, Carlos Bustamante, and Jan Liphardt . Contributed by Carlos Bustamante, December 13, 2006

Prior to this paper the role of light in powering cells containing proteorhodopsin and participation in ocean energy fluxes remained largely unclear. This paper makes an attempt to show that when cellular respiration is inhibited by depleting oxygen or by the respiratory poison azide, Escherichia coli cells expressing PR become light-powered.

Instrumentation

Power density values for green light refer to the power density passed by a D540/25ϫ filter originating at a 175 W Xenon bulb .The sample chamber was periodically illuminated with bright green light (160mW/cm2) coinciding with the maximum of PR’s absorption spectrum, 525 nm. At 525 nm the sample was observed to show maximum absorption (fig.1).
Based on the above fact from the above given reference we decided to use green LED of dominant wavelenght of 525 nm.

Diagrams


fig(1) & fig(2) Copyright@Light-powering Escherichia coli with proteorhodopsin Jessica M. Walter, Derek Greenfield, Carlos Bustamante, and Jan Liphardt. Contributed by Carlos Bustamante, December 13, 2006

Specifications

  1. LM78M05 – 3 terminal positive voltage regulator
  2. LED of wavelength -525 nm( colour- parrot green)
  3. 9 V battery
  4. 100 ohms/51 ohms/ 33 ohms/ 18 ohms resistor

LED:

OVLFx3C7 Series (green): OVLFG3C7(fig.3)

  1. Characteristics
    • High brightness with well-defined spatial radiation patterns
    • UV -resistant epoxy lens
    • Round Through-Hole LED Lamp(5 mm)
  2. Material - InGaN
  3. Typical Intensity - 5200 mcd (millicandela)
  4. Lens colour - water clear
  5. Operating Voltage - (-40 to 85) degree C
  6. Continous forward current - 20mA
  7. Peak Wavelength - 521 nm
  8. Dominant Wavelength - 525 nm
  9. Spectra Half-Width - 25 nm

  10. Circuit Diagram of the Setup


    Observation


    Reading 1 Reading 2 Reading 3 Reading 4
    Vss (V) 5.56 5.56 5.56 5.56
    Vr(V) 0.5 1.0 0.33 0.66
    V led(V) 5.06 4.56 5.23 4.9
    I led(mA) 29.7 26.8 30.7 28.8

    Calculation


    • LED equivalent efficiency - 40 lumen/W
    • Luminous Intensity - 5.2 cd = 65.52 lumens
    • Power given (excluding heat) by 1 LED - 1.638 W
    • Power by 140 LEDs -229.32 W
    • Luminous Intensity -81.105359 mW/cm2
    • Internal Resistance of LED - 3.4 V/ 20 mA = 170 ohms

    Assuming average current across the LED to be 30 mA. This gives an Luminous Efficiency to be 135% (fig.4). So the luminous Intensity of the entire lighting setup = 109.4922 mW/cm2 The 140 LEDs were fixed in 3 breadboards . The luminous intensity calculated above is a cumulative effect of all The LED's. So depending upon how many breadboards we use, we can choose to establish a luminous intensity of 37 mW/cm2, 74 mW/cm2 and 109.4922 mW/cm2. The distance of the lighting source from the flask containing Proteorhodopsin and Retinal was strictly maintained at 15cm.