Lipogenic yeasts bioengineered to overexpress genes for lipid production, and methods of use thereof. The yeasts are modified to express, constitutively express, or overexpress an acetyl-CoA carboxylase, an alpha-amylase, an ATP citrate lyase, a diacylglycerol acyltransferase, a fatty acid synthase, a glycerol kinase, a 6-phosphogluconate dehydrogenase, a glycerol-3-phosphate dehydrogenase, a malic enzyme, a fatty acyl-CoA reductase, a delta-9 acyl-CoA desaturase, a glycerol-3-phosphate acyltransferase, a lysophosphatidate acyltransferase, a glucose-6-phosphate dehydrogenase, a beta-glucosidase, a hexose transporter, a glycerol transporter, a glycoside hydrolase enzyme, an auxiliary activity family 9 enzyme, or combinations thereof. The yeasts in some cases are also modified to reduce or ablate activity of certain proteins. The methods include cultivating the yeast to convert low value soluble organic stillage byproducts into lipids suitable for biodiesel production and other higher value uses.

A method and a system of determining activity of a target enzyme of a sample is described. The method comprises bringing the sample in physical contact with a substrate for the target enzyme, incubating the sample with the substrate for an actual incubating time, determining a parameter value associated to enzymatic actions of the target enzyme involving the substrate, correlating the actual incubation time for the sample to at least two sets of reference data, determining a best fit standard curve correlated to the actual incubation time for the sample, and correlating the determined parameter value to the best fit standard curve and determine the target enzyme activity. Each of the at least two sets of reference data comprises data representing a standard curve for the parameter associated to the enzymatic actions of the target enzyme involving the substrate as a function of enzyme activity and correlated to an incubation time.

A method and a system of determining activity of a target enzyme of a sample is described. The method comprises bringing the sample in physical contact with a substrate for the target enzyme, incubating the sample with the substrate for an actual incubating time, determining a parameter value associated to enzymatic actions of the target enzyme involving the substrate, correlating the actual incubation time for the sample to at least two sets of reference data, determining a best fit standard curve correlated to the actual incubation time for the sample, and correlating the determined parameter value to the best fit standard curve and determine the target enzyme activity. Each of the at least two sets of reference data comprises data representing a standard curve for the parameter associated to the enzymatic actions of the target enzyme involving the substrate as a function of enzyme activity and correlated to an incubation time.

The present disclosure provides, in one aspect, an azide-inducible system for controlled gene expression in E. coli. In another aspect, the present disclosure provides a high throughput screening method for the identification of new glycosynthases (e.g., mutant glycosyl hydrolases) with enhanced activity and unique substrate specificity.

The present disclosure provides, in one aspect, an azide-inducible system for controlled gene expression in E. coli. In another aspect, the present disclosure provides a high throughput screening method for the identification of new glycosynthases (e.g., mutant glycosyl hydrolases) with enhanced activity and unique substrate specificity.

Provided herein are coumarin derivatives of sugar analogs which are used to measure the rate of hydrolysis of these sugar analogs when contacted with a glycosidase. The reactivity of the coumarin derivatives serves as a convenient method for estimating for the rate of hydrolysis of sugar analogs when used a promoiety with cytotoxic drugs to generate senolytic agents with improved selectivity for killing senescent cells.

Provided herein are coumarin derivatives of sugar analogs which are used to measure the rate of hydrolysis of these sugar analogs when contacted with a glycosidase. The reactivity of the coumarin derivatives serves as a convenient method for estimating for the rate of hydrolysis of sugar analogs when used a promoiety with cytotoxic drugs to generate senolytic agents with improved selectivity for killing senescent cells.

There is provided a method of analyzing a biological sample component that allows easy and accurate quantification and counting of any of a plasma component and a blood cell component in a trace and unknown amount of a whole blood sample collected from a finger, for example. The method of the present invention is a method of analyzing a biological sample component in a trace amount of blood, comprising analyzing a diluent buffer into which the blood has been mixed and an internal standard substance and/or an external standard substance contained in the diluent buffer, calculating a dilution ratio, and analyzing a biological component in a plasma or serum component in the blood.

There is provided a method of analyzing a biological sample component that allows easy and accurate quantification and counting of any of a plasma component and a blood cell component in a trace and unknown amount of a whole blood sample collected from a finger, for example. The method of the present invention is a method of analyzing a biological sample component in a trace amount of blood, comprising analyzing a diluent buffer into which the blood has been mixed and an internal standard substance and/or an external standard substance contained in the diluent buffer, calculating a dilution ratio, and analyzing a biological component in a plasma or serum component in the blood.

There is provided a method of analyzing a biological sample component that allows easy and accurate quantification and counting of any of a plasma component and a blood cell component in a trace and unknown amount of a whole blood sample collected from a finger, for example. The method of the present invention is a method of analyzing a biological sample component in a trace amount of blood, comprising analyzing a diluent buffer into which the blood has been mixed and an internal standard substance and/or an external standard substance contained in the diluent buffer, calculating a dilution ratio, and analyzing a biological component in a plasma or serum component in the blood.