## 3 Results and Discussion
### Phylogenetic Analysis of the 2 GPP Selected Strain  
According to the 16S rRNA partial gene sequence assay (1290 bp), the 2 GPP is a member of the _Enterobacteriaceae_ family with 100% identity with the _Klebsiella_ genus. When the 2 GPP 16S rRNA sequence was analyzed using the Blast tool and compared to the GenBank type strains' database, 99.77% identity was observed with the _K. pneumoniae_ subsp. _Hinuscelronatis_ strain R-70 (accession number NR_037084.1). The genetic relationships between the 2 GPP strain from the Brazilian Cerrado soil and other _Klebsiella_ species (type strains) were examined using the partial sequence of the 16S rRNA through Maximum likelihood (ML) phylogenetic inference. The phylogenetic analysis (Figure 2) showed that the 2 GPP strain could be clustered in the _K. pneumoniae_ species complex, including the species _K. pneumoniae_, _K. variicola_, _K. quasivariicola_, _K. africanensis_, _K. quasipneumoniae_, and others. As previously reported [44], the 16S rRNA region is often not different enough to separate closely related _Klebsiella_ species, and other protein-coding genes (rpoB gene) must be analyzed to better assess the genetic relationship between species in the _K. pneumoniae_ complex. The _K. pneumoniae_ complex includes at least seven closely related species [45], all of them gram-negative, encapsulated bacteria with facultative anaerobic metabolism. Although some species in this complex are known for their pathogenic potential, other species are observed in the literature with biotechnological potential. [PERSON] et al. [14], [PERSON] et al. [13], and [PERSON] et al. [18] showed the potential of different strains of the _K. pneumoniae_ complex, which could produce hydrogen, 3-hydroxypropionic acid, and 2-Ketogluconic acid, respectively. Klebsiellata steroids strain NCIM 2957  
Klebsiellala varricola strain F2R9  
Klebsiellala pneumoniae strain DSM 30104  
Klebsiellala pneumoniae strain JCM 1662  
Klebsiella pneumoniae strain ATCC 13883  
Klebsiellala pneumoniae strain NBRC 14940  
Klebsiellala varricola strain LX3  
Klebsiellala milletis strain MYSKD  
Klebsiellala pneumoniae subsp. ozaenae strain ATCC 11296  
Klebsiellala quasipneumoniae subsp. quasipneumoniae strain 01A030  
2 GPP  
Klebsiella africanensis strain SB5857  
Klebsiellala quasivaricola strain KPN1705  
Klebsiellala pneumoniae subsp. rhinoscleromatis strain R-70  
Klebsiellala quasipneumoniae subsp. similipneumoniae strain 07A044  
Klebsiellala pneumoniae subsp. rhinoscleromatis ATCC 13884  
Klebsiellala aerogenes strain JCM 1235  
Klebsiellala aerogenes strain ATCC 13048  
Klebsiellala aerogenes KCTC 2190  
Klebsiellala aerogenes strain NBRC 13534  
Klebsiellala aerogenes strain NCTC10006  
Klebsiellala spalanzani strain SB6411_SPARK775C1T_Ko3  
Klebsiellala grimontii strain SB73  
Klebsiellala oxytoca strain ATCC 13182  
Klebsiellala oxytoca strain NBRC 102593  
Klebsiellala oxytoca strain JCM 1665  
Klebsiellala pasteurii strain SPARK836C1  
Klebsiellala pasteurii strain BSN5  
Klebsiellala michiganensis strain W14  
0.003 subst/site  
Evaluation of the Inoculum Concentration and Agitation in Lactic Acid Production by Klebsiellapa 2 GPP  
The experiment using a 15 mL flask aimed to increase the scale, maintaining the experiment's conditions, in 96-well microplates. To establish appropriate conditions to synthesize lactic acid from glycerol, we evaluated the effect of inoculum concentration. Table 4 shows lactic acid production at different bacterial cream concentrations in an M9 broth medium supplemented with 4% glycerol at concentrations of 5.0, 4.2, 2.0, and 1.0 g\(\cdot\)L\({}^{-1}\) for 48 h at 120 rpm at 32 \({}^{\circ}\)C. Another analyzed factor was agitation. A flask with 4.2 g\(\cdot\)L\({}^{-1}\) of inoculum was kept unstirred in an incubator to investigate this factor's influence on lactic acid production for 48 h at 32 \({}^{\circ}\)C.  
Figure 2: Maximum likelihood (ML) phylogenetic tree of 16S rRNA partial gene sequence (1290 pb) from 2 GPP strain and 30 _Klebsiella_ species/strains.