Production of trypsin MCQs With Answer

Introduction

Production of trypsin MCQs With Answer is designed to help M.Pharm students master concepts of trypsin production, purification, and application in pharmaceutical biotechnology. This set covers zymogen activation, microbial and recombinant expression systems, fermentation parameters, downstream processing including affinity and ion-exchange chromatography, stabilization strategies, immobilization methods, activity assays, inhibition and regulatory considerations. Questions range from fundamentals (trypsinogen activation, molecular weight, pI) to practical production challenges (yield optimization, protease autolysis control, inhibitor management). Targeted for advanced learners, these MCQs emphasize mechanistic understanding and process decision-making used in industrial and research settings.

Q1. Which precursor form of trypsin is secreted by the pancreas and requires activation to become enzymatically active?

• Protrypsin
• Trypsinogen
• Pretrypsin
• Chymotrypsinogen

Correct Answer: Trypsinogen

Q2. Which enzyme is primarily responsible for converting trypsinogen to active trypsin in the small intestine?

• Enterokinase (enteropeptidase)
• Cathepsin D
• Pepsin
• Alkaline phosphatase

Correct Answer: Enterokinase (enteropeptidase)

Q3. Which of the following microbial expression systems is most frequently used for high-yield recombinant trypsin production with minimal glycosylation?

• Pichia pastoris
• Escherichia coli
• CHO cells
• HEK293 cells

Correct Answer: Escherichia coli

Q4. Which chromatography ligand is commonly used for specific affinity purification of trypsin based on its active site interaction?

• Benzamidine-Sepharose
• Glutathione-Sepharose
• Protein A-Sepharose
• Ni-NTA agarose

Correct Answer: Benzamidine-Sepharose

Q5. Which buffer condition helps minimize trypsin autolysis during downstream processing?

• High ionic strength at neutral pH
• Low temperature and slightly acidic pH
• Elevated temperature and alkaline pH
• Presence of calcium chelators like EDTA

Correct Answer: Low temperature and slightly acidic pH

Q6. What is the typical approximate molecular weight of bovine pancreatic trypsin after removal of propeptide?

• 24 kDa
• 54 kDa
• 10 kDa
• 80 kDa

Correct Answer: 24 kDa

Q7. Which assay substrate is commonly used for measuring trypsin activity in enzyme kinetics studies?

• BAEE (Nα-Benzoyl-L-arginine ethyl ester)
• PNPP (p-nitrophenyl phosphate)
• ONPG (o-nitrophenyl-β-D-galactopyranoside)
• DAPG (diacetylphenylyglycine)

Correct Answer: BAEE (Nα-Benzoyl-L-arginine ethyl ester)

Q8. Which naturally occurring inhibitor is commonly used to inhibit trypsin in extraction and assay procedures?

• Soybean trypsin inhibitor (SBTI)
• Alpha-1 antitrypsin
• Corn trypsin inhibitor
• Aprotinin is not used for trypsin

Correct Answer: Soybean trypsin inhibitor (SBTI)

Q9. Immobilization of trypsin on solid supports for industrial use primarily provides which advantage?

• Improved thermal stability and reuse
• Complete elimination of autolysis
• Alteration of cleavage specificity
• Increased molecular weight

Correct Answer: Improved thermal stability and reuse

Q10. During recombinant expression of trypsin in E. coli, why is expression often targeted to inclusion bodies followed by refolding?

• To avoid proteolytic degradation in the cytoplasm
• Because inclusion bodies are catalytically active
• To ensure glycosylation occurs correctly
• To increase native disulfide bond formation in vivo

Correct Answer: To avoid proteolytic degradation in the cytoplasm

Q11. Which additive is commonly added during purification to stabilize trypsin and reduce autoproteolysis by binding calcium?

• Calcium ions (Ca2+)
• EDTA
• Sodium azide
• Urea

Correct Answer: Calcium ions (Ca2+)

Q12. What is the primary reason for activating trypsinogen with enterokinase rather than acid in many recombinant production workflows?

• Specific cleavage at the activation peptide without denaturing the enzyme
• Acid activation is faster and preferred industrially
• Enterokinase adds glycosylation to trypsin
• Enterokinase reduces molecular weight

Correct Answer: Specific cleavage at the activation peptide without denaturing the enzyme

Q13. In gel electrophoresis under reducing SDS-PAGE conditions, what change is expected for trypsin that contains disulfide bonds?

• Migration consistent with monomeric ~24 kDa due to reduction of disulfides
• Formation of higher molecular weight complexes
• Complete degradation to peptides
• No change compared to non-reducing conditions

Correct Answer: Migration consistent with monomeric ~24 kDa due to reduction of disulfides

Q14. Which downstream step is most effective for concentrating crude trypsin solutions without significant activity loss?

• Ultrafiltration with appropriate MWCO membrane
• Lyophilization without any stabilizer
• Dialysis against distilled water overnight
• Heating to precipitate impurities

Correct Answer: Ultrafiltration with appropriate MWCO membrane

Q15. Which parameter is most critical to monitor during fermentation of trypsin-producing recombinant strains to optimize yield?

• Induction timing and inducer concentration
• Color of the culture medium
• Ambient light exposure
• Atmospheric pressure variations

Correct Answer: Induction timing and inducer concentration

Q16. Benzamidine affinity chromatography preferentially interacts with trypsin because benzamidine mimics which of the following?

• Arginine side chain at the substrate P1 position
• Hydrophobic residues at P3 position
• Proline at P2 position
• Glycosylation motifs

Correct Answer: Arginine side chain at the substrate P1 position

Q17. Which storage condition is generally recommended for purified trypsin to retain activity long-term?

• Aliquoted at -20°C or -80°C with glycerol or stabilizer
• Room temperature in buffer without additives
• Repeated freeze-thaw cycles to maintain structure
• Stored dry at 37°C

Correct Answer: Aliquoted at -20°C or -80°C with glycerol or stabilizer

Q18. What is the main regulatory concern for pharmaceutical-grade trypsin used in biologics processing?

• Host-cell protein contamination and viral safety
• Presence of colored dyes
• Ability to activate chymotrypsinogen
• Insolubility in aqueous buffers

Correct Answer: Host-cell protein contamination and viral safety

Q19. Immobilized trypsin reactors used for peptide mapping provide which analytical advantage?

• Short digestion times with reproducible cleavage and easy enzyme removal
• Non-specific cleavage producing diverse fragments
• Permanent covalent modification of peptides
• Inability to reuse the enzyme

Correct Answer: Short digestion times with reproducible cleavage and easy enzyme removal

Q20. Which modification of trypsin would most likely alter its substrate specificity or activity toward basic residues?

• Mutation of the catalytic histidine or serine residues
• Addition of PEG without site-specific attachment
• Concentration by ultrafiltration
• Storage with glycerol

Correct Answer: Mutation of the catalytic histidine or serine residues

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