System for Staged Inoculation of Multiple Cyanobacterial Photobioreactors for Commercial Scale Generation of Cyanobacteria Cultures
AssigneeAlgenol Biofuels Inc.
InventorsBrendan SCOTT, George MEICHEL, Jesse PHILLIPS-KRESS, Jessica BLANKS
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Abstract
A method of rapid simultaneous inoculation of cyanobacteria to multiple commercial-scale closed photobioreactors for the production of a target molecule such as ethanol.
Claims21
A process for inoculating a plurality of closed photobioreactors connected in parallel, each photobioreactor having a volume from about 10 liters to about 100 liters, with a cyanobacteria genetically enhanced for formation of a target molecule, comprising the steps of:
- a. growing an inoculum culture of the genetically enhanced cyanobacteria in a first container to an OD750 of from about 1.0 to about 10.0 in a volume of from about 1 liter to about 10 liters of pre-sterilized medium;
- b. transferring the inoculum culture in a sterile manner under pressure without exposure to atmosphere from the first container to a pre-sterilized second container;
- c. growing the inoculum culture in the second container to an OD750 of from about 1.0 to about 10 in a volume of from about 20 liters to about 100 liters of pre-sterilized medium;
- d. transferring the inoculum culture in a sterile manner under pressure without exposure to atmosphere from the second container to a pre-sterilized third container;
- e. growing the inoculum culture in the third container to an OD750 of from about 2.0 to about 10 in a volume of from about 200 liters to about 1,000 liters of pre-sterilized medium;
- and
- f. transferring the inoculum culture in a sterile manner at a pressure of from about 5 to about 50 psi without exposure to atmosphere from the third container and a sterilized nutrient solution, to a plurality of pre-sterilized photobioreactors comprising a pre-sterilized liquid, wherein said pre-sterilized photobioreactors have inlet tubing comprising a drip emitter and are configured in parallel from the third container, wherein said drip emitter controls the culture flow so that each of the photobioreactors connected in parallel is inoculated at a substantially similar flow rate.
A process for inoculating a plurality of closed photobioreactors connected in parallel, each photobioreactor having a volume from about 10 liters to about 500 liters, with a cyanobacteria genetically enhanced for formation of a target molecule, comprising the steps of:
- a. growing a first inoculum culture of said genetically enhanced cyanobacteria in a first container having a volume of from about 1 liter to about 10 liters to an OD750 of about 1.0 to about 10.0;
- b. transferring in a sterile manner said first inoculum culture under pressure without exposure to atmosphere, to a sterilized second container of a larger size, and adding a volume of sterile medium to create a second inoculum culture of OD750 between 0.04 and 0.5, and growing said second inoculum culture to an OD750 of about 1.0 to about 10;
- and
- c. repeating step b to obtain sufficient volume to fill the plurality of closed photobioreactors.
A process for inoculating a plurality of closed photobioreactors connected in parallel, each photobioreactor having a volume from about 10 liters to about 500 liters, with a cyanobacteria genetically enhanced for formation of a target molecule, comprising the steps of:
- a. determining conditions for growing the enhanced cyanobacteria in the exponential growth phase;
- b. growing a first inoculum culture of said genetically enhanced cyanobacteria in the exponential growth phase in a first container having a volume of from about 1 liter to about 10 liters;
- c. transferring in a sterile manner said first inoculum culture under pressure without exposure to atmosphere, to a sterilized second container of a larger size, in the presence of a volume of sterile medium to create a second inoculum culture of OD750 between 0.04 and 0.5, and growing said second inoculum culture to an OD750 of about 1.0 to about 10;
- d. transferring the second inoculum culture of OD 1 to 10 to a container of larger size comprising sterile liquid to create a third inoculum of OD750 between 0.04 and 0.5 and growing the third inoculum to an OD750 of about 1.0 to about 10;
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- e. transferring the third inoculum culture to each of a plurality of closed photobioreactors connected in parallel;
- wherein the enhanced cyanobacteria form a target molecule within the closed photobioreactors.
Dependencies are inferred from the claim text.