Unit 9: Microbial biotechnology I - Practice Quiz

BTY555 — Biotechnology Laboratory-I 60 Questions
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1 What is the primary function of the enzyme cellulase?

Isolation of cellulases producer microorganisms from soil Easy
A. Breakdown of proteins
B. Breakdown of starch
C. Breakdown of cellulose
D. Breakdown of lipids

2 Cellulose is composed of repeating units of which sugar?

Isolation of cellulases producer microorganisms from soil Easy
A. Fructose
B. Ribose
C. Galactose
D. Glucose

3 Which natural environment is a rich source of cellulose-degrading microorganisms?

Isolation of cellulases producer microorganisms from soil Easy
A. Soil
B. Air
C. Seawater
D. Distilled water

4 Which carbon source is commonly used in media to isolate cellulase producers?

Isolation of cellulases producer microorganisms from soil Easy
A. Glucose
B. Sucrose
C. Glycerol
D. Carboxymethyl cellulose (CMC)

5 Which dye is commonly used to detect cellulase activity on CMC agar plates?

Isolation of cellulases producer microorganisms from soil Easy
A. Safranin
B. Methylene blue
C. Congo red
D. Crystal violet

6 The clear zone around a colony on a Congo red stained CMC plate indicates:

Isolation of cellulases producer microorganisms from soil Easy
A. Spore formation
B. No growth
C. Cellulase production
D. Antibiotic production

7 Which technique is commonly used to obtain isolated colonies from a soil sample?

Isolation of cellulases producer microorganisms from soil Easy
A. Gram staining
B. Centrifugation only
C. Serial dilution and plating
D. Gel electrophoresis

8 Why is serial dilution performed before plating a soil sample?

Isolation of cellulases producer microorganisms from soil Easy
A. To add nutrients
B. To increase cell density
C. To reduce cell density for countable colonies
D. To sterilize the sample

9 A microorganism that can degrade cellulose is termed:

Isolation of cellulases producer microorganisms from soil Easy
A. Proteolytic
B. Amylolytic
C. Lipolytic
D. Cellulolytic

10 Which of the following is a well-known fungal producer of cellulase?

Isolation of cellulases producer microorganisms from soil Easy
A. Lactobacillus
B. Escherichia coli
C. Trichoderma reesei
D. Staphylococcus aureus

11 The medium used to isolate only cellulase producers is best described as:

Isolation of cellulases producer microorganisms from soil Easy
A. Complete medium
B. Enrichment-free medium
C. Selective medium
D. Transport medium

12 During screening, cellulase activity is often visualized by flooding plates with Congo red followed by washing with:

Isolation of cellulases producer microorganisms from soil Easy
A. Acetone
B. Glucose solution
C. Ethanol
D. NaCl solution

13 Cellulose is the main structural component of which of the following?

Isolation of cellulases producer microorganisms from soil Easy
A. Bacterial ribosomes
B. Animal cell membranes
C. Plant cell walls
D. Fungal nuclei

14 Which product is released when cellulase completely hydrolyzes cellulose?

Isolation of cellulases producer microorganisms from soil Easy
A. Nucleotides
B. Fatty acids
C. Amino acids
D. Glucose

15 For measuring cellulase activity, the ratio of clear zone diameter to colony diameter is called the:

Isolation of cellulases producer microorganisms from soil Easy
A. Hydrolysis capacity (index)
B. Growth rate
C. Colony count
D. Dilution factor

16 Before isolating microbes, the glassware and media used must be:

Isolation of cellulases producer microorganisms from soil Easy
A. Dried in sunlight
B. Sterilized
C. Frozen
D. Painted

17 Which equipment is commonly used to sterilize media by moist heat?

Isolation of cellulases producer microorganisms from soil Easy
A. Incubator
B. Autoclave
C. Centrifuge
D. Spectrophotometer

18 After plating, inoculated plates are kept in an incubator mainly to provide:

Isolation of cellulases producer microorganisms from soil Easy
A. Cold shock
B. Light for photosynthesis
C. Optimum temperature for growth
D. Vacuum conditions

19 Which bacterial genus is a common cellulase producer isolated from soil?

Isolation of cellulases producer microorganisms from soil Easy
A. Vibrio
B. Rhizobium
C. Salmonella
D. Bacillus

20 Cellulases have important industrial applications in which of the following?

Isolation of cellulases producer microorganisms from soil Easy
A. Steel manufacturing
B. Biofuel and paper industries
C. Glass cutting
D. Cement production

21 A researcher wants to isolate cellulase-producing microbes from a soil sample. Which soil source is most likely to yield a high number of cellulolytic organisms?

Isolation of cellulases producer microorganisms from soil Medium
A. Deep desert sand with no vegetation
B. Forest litter and decaying plant compost
C. Chlorinated swimming pool water sediment
D. Sterilized potting mix from a sealed bag

22 Carboxymethyl cellulose (CMC) agar is commonly used as a selective medium in cellulase screening. What is the primary role of CMC in this medium?

Isolation of cellulases producer microorganisms from soil Medium
A. It serves as the sole carbon source, favoring cellulose degraders
B. It acts as a pH indicator during incubation
C. It provides nitrogen for microbial growth
D. It functions as a solidifying agent replacing agar

23 After growing colonies on CMC agar, the plate is flooded with Congo red and then washed with 1 M NaCl. What does a clear zone (halo) around a colony indicate?

Isolation of cellulases producer microorganisms from soil Medium
A. Loss of viability of the colony after staining
B. Production of acid that dissolved the agar
C. Hydrolysis of cellulose by secreted cellulase enzymes
D. Accumulation of bacterial pigment in the medium

24 Two isolates form clear zones on CMC agar. Isolate A has a colony diameter of 5 mm with a halo diameter of 20 mm; Isolate B has a colony of 8 mm with a halo of 16 mm. Which isolate has the higher hydrolysis capacity (HC) value?

Isolation of cellulases producer microorganisms from soil Medium
A. Isolate B, with HC = 2.0
B. Both are equal at HC = 2.5
C. Isolate B, with HC = 8.0
D. Isolate A, with HC = 4.0

25 The serial dilution technique is used before plating soil suspensions. What is the main purpose of performing serial dilutions?

Isolation of cellulases producer microorganisms from soil Medium
A. To sterilize the soil sample before plating
B. To obtain well-separated, isolated colonies for screening
C. To increase the total number of microbes present
D. To supply extra nutrients to slow-growing microbes

26 During enrichment culture for cellulase producers, filter paper strips are added to a liquid mineral medium instead of glucose. Why is filter paper preferred here?

Isolation of cellulases producer microorganisms from soil Medium
A. It rapidly lowers the medium pH to inhibit fungi
B. It is nearly pure cellulose, selecting for cellulolytic organisms
C. It prevents oxygen from entering the medium
D. It provides both nitrogen and phosphorus to the culture

27 A student observes that filter paper strips in an enrichment flask become soft, frayed, and eventually disintegrate over several days. What does this observation best suggest?

Isolation of cellulases producer microorganisms from soil Medium
A. Precipitation of mineral salts onto the paper
B. Contamination by non-cellulolytic yeasts only
C. Active cellulase production degrading the filter paper
D. Simple physical soaking with no microbial activity

28 Which combination of enzymes constitutes the complete cellulase system needed for full cellulose degradation by isolated microbes?

Isolation of cellulases producer microorganisms from soil Medium
A. Amylase, lipase, and protease
B. Catalase, oxidase, and urease
C. Endoglucanase, exoglucanase (cellobiohydrolase), and -glucosidase
D. Pectinase, xylanase, and ligninase

29 When preparing a soil suspension for isolation, why is the soil typically mixed with sterile saline (0.85% NaCl) rather than distilled water?

Isolation of cellulases producer microorganisms from soil Medium
A. Saline provides the carbon source for cellulase induction
B. Saline maintains osmotic balance and preserves cell viability
C. Distilled water dissolves cellulose in the soil
D. Saline instantly kills all fungal contaminants

30 A soil sample is diluted to and 0.1 mL is spread on a plate, yielding 48 colonies. What is the estimated cell count per gram of the original soil?

Isolation of cellulases producer microorganisms from soil Medium
A. CFU/g
B. CFU/g
C. CFU/g
D. CFU/g

31 Why is Congo red staining preferred over simply observing colony growth on CMC agar when screening for cellulase producers?

Isolation of cellulases producer microorganisms from soil Medium
A. It measures the exact enzyme concentration in units
B. It visualizes actual cellulose hydrolysis, not just growth
C. It selectively kills non-cellulolytic bacteria on the plate
D. It permanently fixes colonies for long-term storage

32 Which genus is a well-known fungal cellulase producer frequently isolated from soil in laboratory studies?

Isolation of cellulases producer microorganisms from soil Medium
A. Nitrosomonas
B. Trichoderma
C. Rhizobium
D. Lactobacillus

33 To confirm and purify a promising cellulase-producing isolate showing a clear halo, the next logical step is to:

Isolation of cellulases producer microorganisms from soil Medium
A. Add antibiotics to increase enzyme yield
B. Immediately store the mixed plate at C
C. Autoclave the colony to test heat stability of the halo
D. Streak the colony onto fresh CMC agar to obtain a pure culture

34 A quantitative cellulase assay using the DNS (3,5-dinitrosalicylic acid) method measures which product to estimate enzyme activity?

Isolation of cellulases producer microorganisms from soil Medium
A. Ammonia produced during protein breakdown
B. Carbon dioxide evolved during respiration
C. Total protein content of the culture supernatant
D. Reducing sugars released from cellulose hydrolysis

35 Why might a screening plate incubated with glucose-containing CMC agar give misleading results for cellulase detection?

Isolation of cellulases producer microorganisms from soil Medium
A. Glucose lets non-cellulolytic microbes grow and repress cellulase synthesis
B. Glucose converts cellulase into an inactive form permanently
C. Glucose reacts with Congo red to form a false halo
D. Glucose evaporates during incubation, drying the agar

36 The spread plate method is used to distribute a diluted soil sample evenly over the agar surface, which is the reason why this technique is well-suited for isolating aerobic cellulase producers that require good oxygen exposure and surface growth to form measurable clearing zones. Which statement about the pour plate method, in contrast, is correct?

Isolation of cellulases producer microorganisms from soil Medium
A. It requires no molten agar at any stage
B. It cannot produce isolated colonies under any conditions
C. Colonies form only a thin film on top of the agar
D. Colonies grow both within and on the surface of the agar

37 For isolating thermophilic cellulase producers from compost soil, incubation is best carried out at:

Isolation of cellulases producer microorganisms from soil Medium
A. –C to favor psychrophiles
B. –C near boiling temperature
C. –C to favor thermophilic growth
D. –C to favor mesophiles

38 Which control should be included on a CMC-Congo red screening plate to validate that observed halos are due to enzymatic hydrolysis?

Isolation of cellulases producer microorganisms from soil Medium
A. A CMC plate with no inoculum (uninoculated control)
B. A plate incubated without any agar
C. A plate kept in the dark without staining
D. A plate with double the Congo red concentration

39 A soil isolate grows on CMC agar but shows no clear zone after Congo red staining. The most reasonable interpretation is that the isolate:

Isolation of cellulases producer microorganisms from soil Medium
A. Is a strong cellulase producer with a large halo
B. Grows using trace nutrients but does not produce cellulase
C. Cannot survive on any laboratory medium
D. Has completely degraded the agar into liquid

40 When storing a confirmed cellulase-producing bacterial isolate for long-term preservation, which method best maintains viability and genetic stability?

Isolation of cellulases producer microorganisms from soil Medium
A. Leaving the plate on the bench at room temperature
B. Drying the culture in open air on filter paper
C. Repeated daily subculturing on fresh agar
D. Glycerol stock stored at C

41 During screening on carboxymethyl cellulose (CMC) agar, two isolates produce identical clear zone diameters after Congo red staining, but Isolate A shows a much smaller colony diameter than Isolate B. Which conclusion about relative cellulolytic efficiency is most defensible?

Isolation of cellulases producer microorganisms from soil Hard
A. Isolate A has a higher enzymatic index (EI), indicating greater cellulolytic potential per unit biomass
B. Neither can be compared without measuring dry cell weight in liquid culture first
C. Both isolates are equally efficient because clear zone size is the only valid metric
D. Isolate B is more efficient because larger colonies always secrete more enzyme

42 Congo red binds to -1,4-glucan polymers but not to their hydrolysis products. Why does this property make it suitable for visualizing cellulase activity on CMC plates?

Isolation of cellulases producer microorganisms from soil Hard
A. Congo red fluoresces only when bound to glucose monomers released by cellulases
B. Congo red is metabolized by cellulase producers, decolorizing the whole plate uniformly
C. Regions where cellulose is hydrolyzed lose dye binding, producing clear halos against a red background
D. Congo red precipitates intact CMC, creating dark zones over active colonies

43 A researcher performs serial dilution of a soil suspension and plates from the dilution, obtaining 148 colonies. What is the CFU per gram of the original soil sample?

Isolation of cellulases producer microorganisms from soil Hard
A.
B.
C.
D.

44 An enrichment culture uses cellulose as the sole carbon source. After several transfers, a fast-growing non-cellulolytic organism dominates the culture. What is the most likely explanation?

Isolation of cellulases producer microorganisms from soil Hard
A. Enrichment media inherently select against all true cellulose degraders
B. Cross-feeding on soluble sugars released by cellulolytic organisms allows the non-degrader to outcompete producers
C. Cellulose became a preferred carbon source only for the non-degrader over time
D. The non-cellulolytic organism spontaneously acquired cellulase genes during enrichment

45 Why is CMC (a soluble cellulose derivative) preferred over microcrystalline cellulose (Avicel) for primary plate screening of cellulase producers?

Isolation of cellulases producer microorganisms from soil Hard
A. CMC selectively induces only exoglucanases, giving cleaner results
B. Avicel cannot be hydrolyzed by any microbial cellulase system
C. CMC provides more nitrogen, promoting faster colony growth
D. CMC is soluble and yields transparent agar, allowing clear halos to be visualized after dye staining

46 A colony hydrolyzes CMC (endoglucanase-positive) but shows no activity on filter paper. What does this most likely indicate about its cellulase system?

Isolation of cellulases producer microorganisms from soil Hard
A. It lacks the complete synergistic complex needed for crystalline cellulose degradation
B. It cannot secrete any extracellular enzymes
C. It produces only -glucosidase and no endoglucanase
D. It is not a true cellulose degrader under any condition

47 In a DNS (dinitrosalicylic acid) assay for cellulase activity, one unit is defined as the enzyme releasing of reducing sugar per minute. If enzyme releases glucose equivalents in a 30-minute reaction, what is the activity in U/mL?

Isolation of cellulases producer microorganisms from soil Hard
A.
B.
C.
D.

48 Soil samples from a forest floor rich in decaying leaf litter are chosen preferentially for cellulase producer isolation. What is the ecological rationale?

Isolation of cellulases producer microorganisms from soil Hard
A. Decaying litter is sterile and free of competing organisms
B. Forest soils lack nitrogen, forcing all microbes to degrade cellulose
C. Leaf litter raises soil pH, which is essential for cellulase gene expression
D. High lignocellulosic content selectively enriches for cellulolytic microbial communities

49 A student stains a CMC plate with Congo red for 15 minutes, then observes weak, poorly defined halos. Adding a counterstain wash sharpens the zones dramatically. Why?

Isolation of cellulases producer microorganisms from soil Hard
A. kills residual bacteria, revealing true hydrolysis zones
B. chemically hydrolyzes remaining CMC, enlarging the halos
C. removes loosely bound dye and stabilizes the Congo red–cellulose complex, enhancing contrast
D. reacts with Congo red to form a fluorescent product

50 Two isolates have enzymatic indices of and on CMC plates, yet secondary DNS liquid assays show markedly different total cellulase activities. What best explains this discrepancy?

Isolation of cellulases producer microorganisms from soil Hard
A. Enzymatic index and liquid activity are unrelated by definition
B. One isolate contaminated the other during the liquid assay
C. The DNS assay is inherently unreliable and should be ignored
D. Plate halo size reflects enzyme diffusion and endoglucanase, not overall quantitative secretion or full complex activity

51 In an isolation medium, the sole nitrogen source is and the sole carbon source is cellulose. If cellulose degradation is slow, which limiting factor most directly restricts cellulase producer growth?

Isolation of cellulases producer microorganisms from soil Hard
A. Sulfate toxicity prevents cellulase secretion
B. Nitrogen limitation halts all enzyme synthesis immediately
C. Excess ammonium represses ribosome assembly
D. Availability of assimilable carbon from cellulose hydrolysis governs the growth rate

52 A dense soil suspension is plated without adequate dilution, producing confluent growth and merged halos on CMC agar. What is the most rigorous corrective step for accurate screening?

Isolation of cellulases producer microorganisms from soil Hard
A. Measure the largest merged halo and assign it to all colonies
B. Re-plate from higher dilutions to obtain well-separated colonies before halo measurement
C. Increase the Congo red concentration to separate the halos visually
D. Add more agar to slow diffusion and prevent halo merging

53 Why might a genuinely cellulolytic fungal isolate produce a large clear zone yet grow poorly when cellulose is the sole carbon source, whereas a bacterial isolate grows robustly?

Isolation of cellulases producer microorganisms from soil Hard
A. Clear zones are unrelated to enzyme production in fungi
B. Bacteria always outgrow fungi regardless of enzyme profile
C. The fungus may secrete abundant endoglucanase but lack sufficient -glucosidase, causing cellobiose accumulation and feedback inhibition that limits its own carbon uptake
D. Fungi cannot use any sugar released from cellulose

54 When calculating cellulase specific activity, which expression is correct?

Isolation of cellulases producer microorganisms from soil Hard
A.
B.
C.
D.

55 A soil suspension is heat-treated at for 15 minutes before plating for cellulase producers. What is the primary intent, and what is a key limitation?

Isolation of cellulases producer microorganisms from soil Hard
A. It enriches for spore-forming (e.g., thermotolerant Bacillus) cellulolytic bacteria but excludes non-sporulating and many fungal producers
B. It sterilizes the sample completely so no organisms grow
C. It denatures cellulose, improving its degradability
D. It selectively enriches all Gram-negative cellulose degraders

56 In a DNS assay, absorbance is read at against a glucose standard curve with slope and intercept . If sample absorbance is , the reducing sugar concentration is best given by:

Isolation of cellulases producer microorganisms from soil Hard
A.
B.
C.
D.

57 A newly isolated strain shows a large halo on CMC agar but the halo does not increase after 48 h even though the colony keeps growing. What is the most likely interpretation?

Isolation of cellulases producer microorganisms from soil Hard
A. The organism has permanently lost its cellulase genes
B. Continued growth always guarantees continued halo enlargement
C. Enzyme secretion or induction has plateaued, or substrate near the colony is depleted, so halo expansion stops despite continued growth
D. Congo red has completely degraded, so no further zones can appear

58 Which combination of controls is essential to validate that a clear zone on a Congo red CMC plate is genuinely due to enzymatic cellulose hydrolysis?

Isolation of cellulases producer microorganisms from soil Hard
A. Two plates with different agar brands but no organisms
B. Only an uninoculated plate stained with water
C. A single plate observed at two different temperatures
D. An uninoculated CMC plate and a plate inoculated with a known non-cellulolytic strain, both stained identically

59 During optimization, a producer's cellulase activity drops sharply when glucose is added to the medium alongside cellulose. What regulatory phenomenon best explains this?

Isolation of cellulases producer microorganisms from soil Hard
A. Competitive inhibition of -glucosidase by cellulose
B. Substrate saturation of the cellulase active site by glucose
C. Catabolite repression, where readily usable glucose suppresses cellulase gene expression
D. Irreversible thermal denaturation of cellulase by glucose

60 Comparing two isolates: Isolate X gives zone/colony diameters of and Isolate Y gives . Based on enzymatic index, which is the better candidate and why?

Isolation of cellulases producer microorganisms from soil Hard
A. Both are identical because both have zones above
B. Isolate Y, because its absolute zone () is larger
C. Isolate Y, because a larger colony indicates more enzyme
D. Isolate X, because its exceeds Isolate Y's