In biotechnology, agriculture, and pharmaceutical industries, the decomposition of cell walls is a crucial step in extracting active ingredients, preparing protoplasts, or conducting genetic operations. As two efficient cell wall-degrading enzymes, lyticase and snailase have become "star tools" in scientific research and industrial fields due to their specific mechanisms of action and wide range of application scenarios. This article will deeply analyze the action principles, application scenarios, and usage specifications of both, helping you accurately master the practical skills of these two enzymes.
I. Lyticase: "Precise Disassembly Expert" for Fungal Cell Walls
1. Definition and Source
Lyticase is the abbreviation of fungal cell wall lytic enzyme, derived from Arthrobacter luteus. It is a complex hydrolase, mainly composed of β-1,3-glucanase and alkaline phosphatase, which can specifically hydrolyze the β-1,3-glucan structure in the cell walls of yeast and most fungi.
2. Mechanism of Action
Lyticase destroys the integrity of the cell wall by decomposing components such as poly-β(1→3)-glucose in the fungal cell wall, thereby releasing active protoplasts. Its activity unit is defined as: the amount of enzyme required to cause an increase in absorbance (ΔA800) by 0.001 per minute at 25°C and pH 7.5, with a specific activity of ≥200 units/mg.
3. Application Scenarios
3.1 Agricultural field: Deep processing of edible fungi and fungal control, etc.
3.2 Biotechnology field
1) Genomic research: Efficiently lyse yeast cell walls to extract high-quality genomic DNA or plasmid DNA.
2) Protoplast preparation: In yeast transformation and cell fusion experiments, by treating at 30°C for 1 hour and stabilizing osmotic pressure with sorbitol, intact and highly active protoplasts can be obtained.
3) Industrial fermentation: Disrupt yeast cells to improve the extraction efficiency of metabolites (such as recombinant proteins).
4. Usage Guide-Protoplast Preparation Protocol (Spheroplasting protocol)
4.1 Resuspend yeast cells with buffer (1.0 M Sorbitol, 10 mM PIPES, pH 6.5), add 4.0 mL buffer per gram of wet cell weight;
4.2 Centrifuge at 5000 rpm for 5 minutes at room temperature;
4.3 Resuspend the cells again with buffer (4.0 mL per gram of wet cell weight), and add lyticase (50 U per gram of wet cell weight); when using this enzyme initially, it is best to optimize the dosage according to actual conditions.
4.4 Incubate the mixture with gentle shaking at 30°C for 30 minutes, and monitor the formation of protoplasts in the following way:
1) Add 1 μL of sample to 20 μL of buffer, and the protoplasts should remain intact;
2) Add 1 μL of sample to 20 μL of deionized water, and the protoplasts should rupture; observe and compare the two samples under a microscope.
3) Generally, after 45-60 minutes of reaction, the formation rate of protoplasts is >90%. At this time, collect the cells by centrifugation at 4°C, 5000 rpm for 5 minutes;
4) Resuspend the protoplasts again with buffer (2 mL per gram of wet cell weight), and centrifuge at 5000 rpm for 5 minutes.
II. Snailase: "All-Round Cell Wall Breaking Tool" with Multi-Enzyme Synergy
1. Definition and Source
Snailase is a mixed enzyme preparation extracted from the digestive glands (crop and digestive tract) of snails, containing more than 20 kinds of enzymes such as cellulase, pectinase, amylase, and protease, and is a natural multi-enzyme complex system.
2. Mechanism of Action
Snailase achieves efficient cell wall breaking by the synergistic effect of various enzymes to decompose complex components (such as cellulose, pectin, chitin) in the cell walls of plants, fungi, and insects. Its cell wall breaking rate can reach more than 90%, especially suitable for the treatment of yeast cells.
3. Application Scenarios
3.1 Agricultural field
Feed additive: Decompose plant cell walls to improve the digestibility of nutrients in feed.
Food processing: Used in processes such as juice clarification (decomposing pectin), orange removal, and jam making.
3.2 Cosmetics field
According to current known research, pectinase in snailase can reduce scar hyperplasia, chronic ulcers, bedsores, etc.; cellulase can remove the cell walls of eukaryotes. These effects are treasures in the skincare industry. At the same time, snailase can effectively activate skin activity, remove the cuticle on the skin surface and perform deep cleaning, improve the absorption and utilization rate of nutrients, and reduce the skin's long-term dependence on hormones.
3.3 Biotechnology field
Protoplast preparation: After yeast cells are pretreated with sulfhydryl compounds (such as sodium thioacetate), suspend them in sorbitol solution (pH 5.8-7.2), add 30-40 mg of snailase per gram of cells, and incubate at 37°C for 1 hour to obtain highly active protoplasts.
Genetic engineering: Used as a tool enzyme for cell wall removal to assist gene introduction operations.
4. Usage Guide
Concentration and Conditions:
After yeast cells are treated with sulfhydryl compounds, suspend them in isotonic sorbitol solution with pH 5.8-7.2. 30-40 mg of enzyme per gram of cells is incubated at 37°C for 1 hour to dissolve the cell wall, with a breaking rate of over 90%. After enzymatic hydrolysis, centrifugation (600×g, 5 minutes) is required to remove cell debris.
III. Lyticase vs Snailase: How to Choose?
|
Characteristics |
Lyticase |
Snailase |
|
Source |
Arthrobacter luteus |
Snail digestive glands |
|
Enzyme system composition |
Mainly β-1,3-glucanase |
Multi-enzyme complex (cellulase, pectinase, etc.) |
|
Optimal target |
Yeast, most fungi |
Yeast, plant, insect cells |
|
Application focus |
Genomic research, industrial fermentation |
Genomic research, feed, food, medicine and health care |
|
Storage conditions |
-20°C frozen |
4°C dry |
Ordering Information
|
Product Name |
Product Number |
|
Lysozyme |
10402ES |
|
10412ES |
|
|
10406ES |
|
|
Snailase |
10404ES |
|
Lyticase (10 U/μL) |
10403ES |