FYUG Even Semester Exam 2025
BTCDSC-252: Bioprocess Technology

Subject: Biotechnology | Semester: 4th Semester
Full Marks: 70 | Pass Marks: 28 | Time: 3 Hours

UNIT-I

Question 1(a): Define bioprocess technology. 2 Marks

Bioprocess technology is the application of engineering and biological principles to transform biological materials (like cells or enzymes) into useful products, such as pharmaceuticals, food, and biofuels. It integrates biology and chemical engineering to create efficient large-scale production systems.

Question 1(b): What is fed-batch culture? 2 Marks

A fed-batch culture is a biotechnological production process where one or more nutrients are added to the bioreactor during the cultivation. Unlike a standard batch culture, the product remains in the vessel until the end of the run, allowing for higher cell densities and product yields.

Question 1(c): What is continuous culture? 2 Marks

Continuous culture is a system where fresh medium is added continuously to the bioreactor, while an equal volume of culture (including cells and waste products) is simultaneously removed. This maintains the microbial population in a steady-state exponential growth phase.

Question 2(a): Write a detailed account on basic principle and components of fermentation technology. Add a note on applications of bioprocess technology. 7+3=10 Marks

Principle of Fermentation Technology:

The core principle involves the controlled growth of microorganisms (or animal/plant cells) in a nutrient medium to produce secondary metabolites or biomass. It relies on providing an optimized environment—temperature, pH, aeration, and nutrient supply—to maximize the biocatalytic activity of the organisms.

Basic Components of Fermentation:

  • Inoculum: A healthy, pure culture of the microorganism used to start the fermentation.
  • Culture Media: Nutrient source providing carbon, nitrogen, minerals, and growth factors.
  • Bioreactor (Fermenter): The vessel where the reaction occurs, equipped with control systems.
  • Sterilization System: Ensures the media and vessel are free from contaminants.
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  • Downstream Processing: Steps to recover and purify the final product[span_14](end_span).

Applications of Bioprocess Technology:

  • Healthcare: Production of antibiotics, vaccines, and insulin.
  • Environment: Waste treatment and bioremediation of pollutants.
  • Food Industry: Production of amino acids, organic acids, and alcoholic beverages.
  • Energy: Development of biofuels like bio-ethanol and biodiesel.

Question 2(b): Write notes on: (i) Batch culture (ii) Phases of growth 5+5=10 Marks

(i) Batch culture:

In a batch culture, a fixed volume of medium is inoculated with a microorganism in a closed system. No additional nutrients are added after the start, and nothing is removed until the end of the process, except for air and gases.

(ii) Phases of growth:

Microbial growth in a batch system follows four distinct phases:

  1. Lag Phase: Cells adapt to the new environment; no increase in number.
  2. Log (Exponential) Phase: Cells divide at a constant, maximum rate.
  3. Stationary Phase: Growth rate equals death rate due to nutrient depletion or waste accumulation.
  4. Death (Decline) Phase: Cells die exponentially as conditions become toxic.

UNIT-II

Question 3(a): Define bioreactor. 2 Marks

A bioreactor is a specialized vessel or container that provides a controlled environment for the growth of biological organisms and the conversion of raw materials into specific products via biochemical reactions.

Question 3(b): What is the function of agitator and sparger in a bioreactor? 2 Marks

  • Agitator (Impeller): Mixes the culture to ensure uniform distribution of nutrients and temperature while breaking up air bubbles.
  • Sparger: Introduces sterile air or oxygen into the bioreactor liquid in the form of small bubbles to facilitate aeration.

Question 3(c): Write a note on packed tower bioreactor. 2 Marks

A packed tower bioreactor is a vessel filled with solid packing material (like plastic rings or ceramic beads) onto which microorganisms are immobilized. The nutrient medium flows over the packing, and air is usually introduced from the bottom. It is commonly used for effluent treatment.

Question 4(a): Give a detailed account of the design and components of a bioprocess vessel. 10 Marks

A standard bioprocess vessel (Stirred Tank Bioreactor) is designed for aseptic operation and effective mass transfer.

Key Design Components:

  • Vessel Material: Usually 316L stainless steel for durability and non-toxicity.
  • Baffles: Metal strips attached to the walls to prevent vortex formation and improve mixing.
  • Cooling Jacket/Coils: To remove metabolic heat and maintain constant temperature.
  • Headspace: Extra volume (usually 20-30%) allowed for foaming and gas exchange.
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  • Ports: Entry/exit points for sensors (pH, DO, Temp), inoculation, and sampling[span_38](end_span).

Question 4(b): Write notes on: (i) Airlift bioreactor (ii) Cyclone column bioreactor 5+5=10 Marks

(i) Airlift bioreactor:

These reactors use air/gas to achieve both aeration and agitation without mechanical moving parts. They consist of a "riser" (where gas is injected) and a "downcomer" (where liquid circulates back), creating a highly efficient loop of circulation.

(ii) Cyclone column bioreactor:

This reactor utilizes centrifugal forces (cyclone action) to separate gas from liquid or to enhance mixing. The medium is often pumped tangentially into the column, creating a thin film or vortex that maximizes the surface area for oxygen transfer.


UNIT-III

Question 5(a): What is upstream processing? 2 Marks

Upstream processing refers to the early stages of a bioprocess, including inoculum development, media formulation, and sterilization, ending just before the actual fermentation begins in the bioreactor.

Question 5(b): What is the basic composition of culture media? 2 Marks

  • Carbon Source: e.g., Glucose, Molasses.
  • Nitrogen Source: e.g., Ammonia salts, Yeast extract.
  • Water: The primary solvent.
  • Minerals & Growth Factors: e.g., P, S, Mg, and vitamins.

Question 5(c): Write a note on pure culture. 2 Marks

A pure culture is a laboratory culture containing only a single species or strain of microorganism, free from any contaminating organisms. Maintaining pure cultures is vital for consistent product quality in industrial fermentation.

Question 6(a): Give a detailed account of sterilization. 10 Marks

Sterilization is the total destruction of all living organisms and spores to ensure monosepsis in the fermentation vessel.

  • Batch Sterilization: The medium is heated in the bioreactor itself (typically 121°C for 20-30 mins).
  • Continuous Sterilization: The medium is passed through a heat exchanger, reaching high temperatures (140°C) for very short durations (seconds), which minimizes nutrient degradation.
  • Filter Sterilization: Used for heat-sensitive components; medium is passed through membranes with 0.22 μm pores to physically remove microbes.

Question 6(b): Write notes on: (i) Inocula development (ii) Media preparation. Add a note on strain improvement. 5+5=10 Marks

(i) Inocula development:

The process of progressively increasing the volume of a pure culture from a master vial (stock) to a large volume sufficient to seed the main production fermenter (typically 5-10% of total volume).

(ii) Media preparation:

Involves the precise formulation and dissolution of nutrients to support the target organism's growth and product synthesis.

Strain Improvement:

Techniques like mutagenesis (using UV or chemicals) and recombinant DNA technology are used to enhance the metabolic efficiency of the organism, making it produce more of the desired product.


UNIT-IV

Question 7(a): Define mass transfer. 2 Marks

Mass transfer is the movement of chemical species from a region of high concentration to a region of low concentration. In bioprocessing, it primarily refers to the transfer of oxygen from gas bubbles into the liquid medium where cells can consume it.

Question 7(b): Write about oxygen requirement in bioprocessing. 2 Marks

Aerobic microorganisms require a continuous supply of dissolved oxygen (DO) for respiration and product synthesis. Since oxygen is poorly soluble in aqueous media, it often becomes the limiting factor in high-density cultures.

Question 7(c): What is kLa? 2 Marks

kLa is the volumetric oxygen mass transfer coefficient. It represents the efficiency of a bioreactor in transferring oxygen from gas to liquid. High kLa values indicate superior aeration performance.

Question 8(a): Give an illustrated account of bioprocess measurement and control system. 10 Marks

A control system maintains optimized conditions within the bioreactor by monitoring variables via sensors and adjusting them via actuators.

  • pH Control: Measured by pH electrodes; maintained by adding acid or base pumps.
  • Temperature Control: Measured by thermistors; maintained by cold water flow in jackets.
  • Dissolved Oxygen (DO): Measured by polarographic probes; maintained by adjusting agitation speed or air flow.
  • Antifoam Control: Probes detect foam; triggers the addition of antifoam agents.

Question 8(b): Write notes on: (i) Factors affecting kLa (ii) Computer-aided process control 5+5=10 Marks

(i) Factors affecting kLa:

  • Agitation Speed: Higher speed breaks bubbles, increasing surface area.
  • Air Flow Rate: Increases the volume of air available.
  • Medium Viscosity: Higher viscosity decreases oxygen diffusion and kLa.
  • Temperature: Affects solubility and diffusion rates.

(ii) Computer-aided process control:

Involves using software (SCADA) to collect real-time data from sensors, perform complex calculations, and execute precise automated adjustments, ensuring high reproducibility and safety.


UNIT-V

Question 9(a): What is product recovery? 2 Marks

Product recovery is the set of operations performed after fermentation to separate the desired biological product from the culture medium, cells, and impurities.

Question 9(b): Write a note on solid-liquid separation. 2 Marks

This is the first step of downstream processing, where the biomass (cells) is separated from the fermentation broth using techniques like centrifugation or filtration.

Question 9(c): Write about industrial production of amylase. 2 Marks

Amylase is produced industrially primarily using Bacillus subtilis or Aspergillus oryzae via submerged fermentation. It is used extensively in the starch, textile, and detergent industries.

Question 10(a): Write a detailed account of downstream processing. Add a note on production of ethanol. 6+4=10 Marks

Downstream Processing (DSP) Steps:

  1. Removal of Insolubles: Centrifugation or filtration to remove cells.
  2. Cell Disruption: Required if the product is intracellular (e.g., using high-pressure homogenizers).
  3. Isolation/Concentration: Reducing volume using evaporation or ultrafiltration.
  4. Purification: Using chromatography (ion-exchange, affinity) to reach high purity.
  5. Polishing: Final steps like crystallization or freeze-drying.

Production of Ethanol:

Produced by the fermentation of sugars (molasses or starch) using the yeast Saccharomyces cerevisiae under anaerobic conditions. The ethanol is then recovered and concentrated using fractional distillation.

Question 10(b): Write notes on: (i) Production of lactic acid (ii) Production of SCP 5+5=10 Marks

(i) Production of lactic acid:

Produced by Lactobacillus species through the fermentation of carbohydrates. The pH must be controlled by adding calcium carbonate, forming calcium lactate, which is later acidified to recover pure lactic acid.

(ii) Production of SCP (Single Cell Protein):

SCP refers to edible protein extracted from pure microbial cultures (algae, fungi, or bacteria). Organisms like Spirulina are grown on large scales, harvested, and dried to be used as high-protein supplements for humans or animals.


Exam Focus & Strategy

Important Concepts List

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  • Upstream vs Downstream: Clear distinction between preparation and recovery[span_89](end_span).
  • kLa: Understand it as the "aeration efficiency" metric.
  • Sterilization: Knowing the difference between batch and continuous methods.

Common Mistakes to Avoid

  • Confusing Fed-batch (adding nutrients) with Continuous (adding and removing).
  • Neglecting to mention sterilization when describing bioreactor design.
  • Forgetting that downstream processing is often the most expensive part of a bioprocess.

Answer Presentation Strategy

  • Always mention specific microorganisms for industrial production questions (e.g., S. cerevisiae for Ethanol).
  • Use flowcharts to describe the stages of Downstream Processing.
  • Draw a labeled diagram for Airlift or Stirred Tank bioreactors.