Table 2.
Muramidase activity of Lys c-1 against various bacteria in lysoplate assays. Mosquito derived bacteria were indentified by BLAST search based on 16s rRNA sequences. Purified Lys c-1 (10 μL of 0.01 mg/mL) was incubated with various bacteria derived from female mosquitoes as well as stock bacteria.
| Bacterial isolates from | GenBank accession no. |
Identification (top matches)++ | Sequence length/query coverage/identity% |
Muramidase activity in lysoplate assays** |
|
|---|---|---|---|---|---|
| An. gambiae G3 | Gram-type | Purified Lys c-1 | |||
| G3-1 | FJ816020 | Elizabethkingia meningoseptica * | 1402/99/99 | Gram− | nd |
| G3-3 | FJ816021 | Asaia krungthepensis | 1361/99/99 | Gram− | nd |
| G3-6 | FJ816022 | Micrococcus luteus | 1387/99/100 | Gram+ | nd |
| An. gambiae 4a rr | |||||
| GS-1 | FJ816023 | Erwinia sp. | 1403/100/100 | Gram− | nd |
| GS-2 | FJ816024 | Serratia sp. | 1350/99/99 | Gram− | nd |
| GS-3 | FJ816025 | Uncultured bacterium | 1385/100/99 | Gram+ | 5.5 |
| GS-4 | FJ816026 | Klebsiella oxytoca | 1400/100/99 | Gram− | tr |
| GS-6 | FJ816027 | Uncultured bacterium | 1411/100/99 | Gram− | nd |
| GS-7 | FJ816028 | Elizabethkingia meningoseptica* | 1374/100/99 | Gram− | nd |
| An. Stephensi | |||||
| ST-2 | FJ816029 | Serratia sp. | 1335/100/99 | Gram− | nd |
| ST-3 | FJ816030 | Bacterium spbL1type1-2 | 1327/100/100 | Gram− | nd |
| Non-mosquito derived bacteria | |||||
| Micrococcus luteus | Gram+ | 6 | |||
| Serratia marcescens | Gram− | nd | |||
| Enterobacter cloacae | Gram− | nd | |||
| Staphylococcus epidermidis | Gram+ | nd | |||
| Bacillus subtilis | Gram+ | 7 | |||
| Bacillus thuringiensis | Gram+ | nd | |||
| Asaia sp. | Gram− | nd | |||
| Pseudomonas aeruginosa | Gram− | nd | |||
| Enterobacter sp. | Gram− | nd | |||
|
Micrococcus lysodeikticus (lyophilized) |
Gram+ | 14 | |||
| Escherichia coli DH5α | Gram− | nd | |||
++
Top score from NCBI BLAST database nr/nt; Query coverage is over the entire length of sequenced PCR product as indicated
*
also known as Chryseobacterium meningosepticum; Zone of clearance measured in mm; Measurements were made after O/N incubation of the bacteria with lysozyme sample; nd-absence of detectable zone of clearance; tr-trace zone of clearance (below 5 mm).