Delta G With kcat and Km Calculator

Estimate delta G from kinetic enzyme inputs. Review efficiency, barriers, ratios, and export results quickly. Use flexible units for reliable enzyme comparisons every time.

Calculator Inputs

Turnover number in s^-1.
Michaelis constant value.
Temperature in Celsius.
Reference kcat/Km in M^-1 s^-1.
Used for saturation and rate estimates.
Used for initial velocity.

Formula Used

Catalytic efficiency: kcat / Km

Relative delta G: ΔG = -RT ln((kcat / Km) / reference efficiency)

Activation estimate: ΔG‡ = RT ln(kBT / h kcat)

Saturation fraction: [S] / (Km + [S])

Initial velocity: v0 = kcat[E][S] / (Km + [S])

R is the gas constant. T is temperature in Kelvin. kB is Boltzmann's constant. h is Planck's constant. The relative delta G result is a comparison estimate. It is not a direct equilibrium free energy measurement.

Example Data Table

Example kcat Km Reference Efficiency Catalytic Efficiency Relative ΔG ΔG‡ Estimate
Standard enzyme 120 s^-1 50 uM 1.00E+6 M^-1 s^-1 2.40E+6 M^-1 s^-1 -2.17 kJ/mol 61.16 kJ/mol
Weak variant 25 s^-1 0.2 mM 1.00E+6 M^-1 s^-1 1.25E+5 M^-1 s^-1 5.15 kJ/mol 65.04 kJ/mol
Improved variant 500 s^-1 10 uM 1.00E+6 M^-1 s^-1 5.00E+7 M^-1 s^-1 -9.70 kJ/mol 57.62 kJ/mol

How To Use This Calculator

  1. Enter kcat in per second.
  2. Enter Km and choose its concentration unit.
  3. Add assay temperature in Celsius.
  4. Enter a reference catalytic efficiency.
  5. Add substrate and enzyme concentrations when rate output is needed.
  6. Choose the preferred energy unit.
  7. Press the calculate button.
  8. Download the result as CSV or PDF.

Delta G From Enzyme Kinetics

Delta G can connect enzyme kinetics with thermodynamic style comparisons. This calculator treats kcat and Km as practical kinetic inputs. It does not prove full reaction equilibrium. Instead, it gives useful apparent free energy values. The main output uses catalytic efficiency, kcat divided by Km. That value shows how strongly turnover and substrate binding work together.

Why This Estimate Matters

Enzyme studies often compare mutants, buffers, pH levels, or temperatures. Raw kcat values can rise while Km also rises. That mixed change may hide the real performance shift. Catalytic efficiency gives one combined number. A relative delta G then converts that ratio into energy language. Negative values suggest improved apparent performance versus the chosen reference. Positive values suggest weaker apparent performance.

The tool also reports an Eyring activation estimate from kcat. This barrier uses temperature and assumes kcat behaves like a rate constant. It is best viewed as a model estimate. It helps compare runs under similar assumptions. It should not replace calorimetry or full equilibrium work.

Inputs You Can Control

Enter kcat in per second. Enter Km in micromolar, millimolar, or molar units. Add temperature because RT changes with heat. Choose a reference catalytic efficiency for comparison. A reference of one is simple. A mutant or wild type value is often better. Add substrate concentration to estimate saturation and observed first order turnover. Add enzyme concentration to estimate initial velocity.

Reading The Results

The efficiency result uses molar units. The relative delta G uses negative RT times the natural log of efficiency over reference efficiency. When the ratio equals one, the relative value is zero. The activation estimate uses RT times the natural log of kBT over h kcat. Lower activation values usually mean faster turnover.

Use results as screening values. Keep units consistent. Compare only experiments measured under similar conditions. Temperature, buffer, isotope effects, and assay design can change interpretation. Export the table when you need records. The CSV file supports spreadsheets. The PDF file supports simple reporting. Good documentation also matters. Store every assumption beside each result. Record units, reference values, and assay notes. Small reporting gaps can make later comparisons confusing, especially during kinetic model review, manuscript preparation, and scientific work.

FAQs

What does this calculator estimate?

It estimates catalytic efficiency, relative delta G, activation delta G, saturation fraction, observed turnover, and initial velocity from kcat, Km, temperature, substrate, and enzyme inputs.

Is this true reaction delta G?

No. It is an apparent comparison based on kinetic efficiency. True reaction delta G requires equilibrium data or related thermodynamic measurements.

Why is Km converted to molar units?

Catalytic efficiency commonly uses M^-1 s^-1. Converting Km to molar units keeps the formula consistent and prevents unit errors.

What reference efficiency should I use?

You can use one for a simple scale. For enzyme comparison, use a wild type, control assay, or published reference efficiency.

What does a negative relative delta G mean?

It means the calculated catalytic efficiency is higher than the reference. In this model, that suggests improved apparent kinetic performance.

What does the activation estimate mean?

It uses an Eyring style equation with kcat. It is a useful model barrier, not a full mechanistic proof.

Can I compare different temperatures?

You can calculate them, but compare carefully. Temperature changes both RT and enzyme behavior, so assay conditions should be documented.

Why include substrate and enzyme concentration?

They allow saturation and initial velocity estimates. These outputs help connect kinetic constants with expected assay performance.

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Important Note: All the Calculators listed in this site are for educational purpose only and we do not guarentee the accuracy of results. Please do consult with other sources as well.