Recently, we proposed a novel and effective strategy for enzyme immobilization, namely, as elastin-like polypeptide (ELP)-mediated silicification-based protein self-immobilization (ESPSI). Here, an enzyme with an acidic isoelectric point (pI) was chosen to further demonstrate the feasibility and generality of this method; pI is a critical factor in biosilicification. The lichenase (BglS) from Bacillus subtilis 168, with a theoretical pI of 5.77, was genetically fused to the cationic ELP. The recombinant chimera protein BglS-ELP (B-E) was purified through ELP-based nonchromatographic inverse transition cycling (ITC) method. B-E then was self-encapsulated within silica nanoparticle (NP) via ELP-mediated biomimetic silicification, producing the nanobiocatalyst B-E@silica. Excellent encapsulation efficiency (>85%) could be achieved within a short immobilization time (10 min). In addition, the encapsulation efficiency could be promoted through temperature-induced phase transition. Meanwhile, this method gave negligible protein leakage (<0.5%). Thus, the B-Epsilica showed good reusability, retaining similar to 80% of the initial activity after 10 reaction cycles. These results indicated that ESPSI was suitable for the protein with an acidic pI. In addition, phase transition was a unique route to improve immobilization efficiency.