Notice of Pre-AIA or AIA Status
The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA .
DETAILED ACTION
This application is a 371 of PCT/EP2022/065632.
The amendment filed on June 8, 2026 has been entered. No new matter has been entered.
Election/Restrictions
Applicant’s election without traverse of Group II with a species election of FCb2 Km82 from Kluyveromyces marxianus (i.e. SEQ ID NO:10) as the recombinant flavocytochrome b2 in the reply filed on June 8, 2026 is acknowledged.
Status of Claims
Claims 1-5 are pending.
Claims 1-5 are under examination.
Foreign Priority
Receipt is acknowledged of certified copies of papers required by 37 CFR 1.55.
Information Disclosure Statement
The information disclosure statement (IDS) submitted on February 29, 2024 and July 16, 2026 are in compliance with the provisions of 37 CFR 1.97. Accordingly, the information disclosure statements are being considered by the examiner.
Nucleotide and/or Amino Acid Sequence Disclosures
REQUIREMENTS FOR PATENT APPLICATIONS CONTAINING NUCLEOTIDE AND/OR AMINO ACID SEQUENCE DISCLOSURES
Items 1) and 2) provide general guidance related to requirements for sequence disclosures.
37 CFR 1.821(c) requires that patent applications which contain disclosures of nucleotide and/or amino acid sequences that fall within the definitions of 37 CFR 1.821(a) must contain a "Sequence Listing," as a separate part of the disclosure, which presents the nucleotide and/or amino acid sequences and associated information using the symbols and format in accordance with the requirements of 37 CFR 1.821 - 1.825. This "Sequence Listing" part of the disclosure may be submitted:
In accordance with 37 CFR 1.821(c)(1) via the USPTO patent electronic filing system (see Section I.1 of the Legal Framework for Patent Electronic System (https://www.uspto.gov/PatentLegalFramework), hereinafter "Legal Framework") as an ASCII text file, together with an incorporation-by-reference of the material in the ASCII text file in a separate paragraph of the specification as required by 37 CFR 1.823(b)(1) identifying:
the name of the ASCII text file;
ii) the date of creation; and
iii) the size of the ASCII text file in bytes;
In accordance with 37 CFR 1.821(c)(1) on read-only optical disc(s) as permitted by 37 CFR 1.52(e)(1)(ii), labeled according to 37 CFR 1.52(e)(5), with an incorporation-by-reference of the material in the ASCII text file according to 37 CFR 1.52(e)(8) and 37 CFR 1.823(b)(1) in a separate paragraph of the specification identifying:
the name of the ASCII text file;
the date of creation; and
the size of the ASCII text file in bytes;
In accordance with 37 CFR 1.821(c)(2) via the USPTO patent electronic filing system as a PDF file (not recommended); or
In accordance with 37 CFR 1.821(c)(3) on physical sheets of paper (not recommended).
When a “Sequence Listing” has been submitted as a PDF file as in 1(c) above (37 CFR 1.821(c)(2)) or on physical sheets of paper as in 1(d) above (37 CFR 1.821(c)(3)), 37 CFR 1.821(e)(1) requires a computer readable form (CRF) of the “Sequence Listing” in accordance with the requirements of 37 CFR 1.824.
If the "Sequence Listing" required by 37 CFR 1.821(c) is filed via the USPTO patent electronic filing system as a PDF, then 37 CFR 1.821(e)(1)(ii) or 1.821(e)(2)(ii) requires submission of a statement that the "Sequence Listing" content of the PDF copy and the CRF copy (the ASCII text file copy) are identical.
If the "Sequence Listing" required by 37 CFR 1.821(c) is filed on paper or read-only optical disc, then 37 CFR 1.821(e)(1)(ii) or 1.821(e)(2)(ii) requires submission of a statement that the "Sequence Listing" content of the paper or read-only optical disc copy and the CRF are identical.
Specific deficiencies and the required response to this Office Action are as follows:
Specific deficiency – Nucleotide and/or amino acid sequences appearing in the specification (page 39) are not identified by sequence identifiers in accordance with 37 CFR 1.821(d).
Required response – Applicant must provide:
A substitute specification in compliance with 37 CFR 1.52, 1.121(b)(3) and 1.125 inserting the required sequence identifiers, consisting of:
A copy of the previously-submitted specification, with deletions shown with strikethrough or brackets and insertions shown with underlining (marked-up version);
A copy of the amended specification without markings (clean version); and
A statement that the substitute specification contains no new matter.
Specific deficiency – Nucleotide and/or amino acid sequences appearing in the drawings are not identified by sequence identifiers in accordance with 37 CFR 1.821(d). Sequence identifiers for nucleotide and/or amino acid sequences must appear either in the drawings or in the Brief Description of the Drawings (Figures 1-7, 8C, 9C, and 10C).
Required response – Applicant must provide:
Replacement and annotated drawings in accordance with 37 CFR 1.121(d) inserting the required sequence identifiers;
AND/OR
A substitute specification in compliance with 37 CFR 1.52, 1.121(b)(3) and 1.125 inserting the required sequence identifiers into the Brief Description of the Drawings, consisting of:
A copy of the previously-submitted specification, with deletions shown with strikethrough or brackets and insertions shown with underlining (marked-up version);
A copy of the amended specification without markings (clean version); and
A statement that the substitute specification contains no new matter.
Specification
The disclosure is objected to because it contains an embedded hyperlink and/or other form of browser-executable code (pages 34-35, 37-38, 51, and 60-62). Applicant is required to delete the embedded hyperlink and/or other form of browser-executable code; references to websites should be limited to the top-level domain name without any prefix such as http:// or other browser-executable code. See MPEP § 608.01.
Claim Rejections - 35 USC § 112
The following is a quotation of 35 U.S.C. 112(b):
(b) CONCLUSION.—The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the inventor or a joint inventor regards as the invention.
The following is a quotation of 35 U.S.C. 112 (pre-AIA ), second paragraph:
The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the applicant regards as his invention.
Claim 2 is rejected under 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph, as being incomplete for omitting essential elements, such omission amounting to a gap between the elements. See MPEP § 2172.01. The omitted elements are: elements i.-iv. Appropriate correction is required.
Claim 4 is rejected under 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph, as being indefinite for failing to particularly point out and distinctly claim the subject matter which the inventor or a joint inventor (or for applications subject to pre-AIA 35 U.S.C. 112, the applicant), regards as the invention.
Claim 4 recites the phrase “electrode of claim 5, having an amino acid sequence selected from …” The metes and bounds of the phrase in the context of the above claim are not clear. It is unclear how an electrode comprises an amino acid sequence. Clarification is requested.
Claim 5 and claims 1-4 depending therefrom are rejected under 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph, as being incomplete for omitting essential elements, such omission amounting to a gap between the elements. See MPEP § 2172.01. The omitted elements are: elements i.-vii. Appropriate correction is required.
Claim 5 and claims 1-4 depending therefore are rejected under 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph, as being indefinite for failing to particularly point out and distinctly claim the subject matter which the inventor or a joint inventor (or for applications subject to pre-AIA 35 U.S.C. 112, the applicant), regards as the invention.
Claim 5 recites the limitation "the length of the sequence of iii." in line 14. There is insufficient antecedent basis for this limitation in the claim. Claim 5 does not recite an “amino acid sequence of iii”. Clarification is requested.
The following is a quotation of the first paragraph of 35 U.S.C. 112(a):
(a) IN GENERAL.—The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor or joint inventor of carrying out the invention.
The following is a quotation of the first paragraph of pre-AIA 35 U.S.C. 112:
The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor of carrying out his invention.
Claims 1, 3, and 5 are rejected under 35 U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA ), first paragraph, as failing to comply with the written description requirement. The claim(s) contains subject matter which was not described in the specification in such a way as to reasonably convey to one skilled in the relevant art that the inventor or a joint inventor, or for pre-AIA the inventor(s), at the time the application was filed, had possession of the claimed invention.
MPEP 2111.01 states that ''[d]uring examination, the claims must be interpreted as broadly as their terms reasonably allow.'' In this case, the examiner has broadly interpreted the claims to encompass an electrode comprising (A) any mature flavocytochrome b2 (FCb2) or mature Kluyveromyces marxianus FCb2 and any functionally active variant thereof, wherein the N-terminus consists of (B)(1) the signal peptide sequence of SEQ DI NO:40, 41, or 42 or (2) any signal peptide sequence having viii. a Met residue, ix. any tag sequence, any affinity tags, any solubility enhancement tags, and any monitoring tags, x. an amino acid sequence of a length of 0-9 the native signal peptide sequence, and xi. I/L/V-xN/A/L motif, which can be of number 0, 1 or partially truncated. Therefore, the claims are drawn to an electrode comprising a genus of FCb2 having unknown structure but having FCb2 activity.
MPEP 2163 I. states that to “satisfy the written description requirement, a patent specification must describe the claimed invention in sufficient detail that one skilled in the art can reasonably conclude that the inventor had possession of the claimed invention.
MPEP 2163. II.A.3.(a) sates that “Possession may be shown in many ways. For example, possession may be shown by describing an actual reduction to practice of the claimed invention. Possession may also be shown by a clear depiction of the invention in detailed drawings or in structural chemical formulas which permit a person skilled in the art to clearly recognize that inventor had possession of the claimed invention. An adequate written description of the invention may be shown by any description of sufficient, relevant, identifying characteristics so long as a person skilled in the art would recognize that the inventor had possession of the claimed invention.
According to MPEP 2163.II.A.3.(a).ii), “Satisfactory disclosure of a ‘representative number’ depends on whether one of skill in the art would recognize that the applicant was in possession of the necessary common attributes or features possessed by the members of the genus in view of the species disclosed. For inventions in an unpredictable art, adequate written description of a genus which embraces widely variant species cannot be achieved by disclosing only one species within the genus…Instead, the disclosure must adequately reflect the structural diversity of the claimed genus, either through the disclosure of sufficient species that are ‘representative of the full variety or scope of the genus,’ or by the establishment of ‘a reasonable structure-function correlation.’"
The recitation of “flavocytochrome b2” and “signal peptide sequence” fails to provide a sufficient description of the genus of the polypeptides as it merely describes the functional features of the genus without providing any definition of the structural features of the species within the genus. The specification does not specifically define any of the species that fall within the genus. The specification does not define any structural features commonly possessed by members of the genus that distinguish them from others. One skilled in the art therefore cannot, as one can do with a fully described genus, visualize or recognize the identity of the members of the genus.
W0T5A6 (UnitProtKB/TrEMBL Database. February 26, 2020 – form PTO-892) discloses a Kluyveromyces marxianus flavocytochrome b2 having 100% sequence identity to the flavocytochrome b2 of SEQ ID NO:7 of the instant application (pages 1-2 and see the sequence alignment below). W0T5A6 discloses that the amino acids 89-166 is the cytochrome heme binding domain and amino acids 198-565 is the FMN domain (page 2). However, the prior art does not teach an electrode comprising any truncated flavocytochrome b2 comprising any signal peptide and/or tag sequence at the N-terminus.
The specification is limited to description of an electrode comprising a recombinant flavocytochrome having the amino acid sequence of SEQ ID NO:8, 9, or 10, which is composed of the signal peptide of SEQ ID NO:40, 41, or 42, respectively, and mature Kluyveromyces marxianus flavocytochrome b2. While MPEP 2163 acknowledges that in certain situations “one species adequately supports a genus,” it also acknowledges that “[f]or inventions in an unpredictable art, adequate written description of a genus which embraces widely variant species cannot be achieved by disclosing only one species within the genus.” In view of the widely variant species encompassed by the genus, the above examples described above are not enough and does not constitute a representative number of species to describe the whole genus. Therefore, the specification fails to describe a representative species of the claimed genus.
Further, one of skill in the art could identify variants of a flavocytochrome b2, Kluyveromyces marxianus flavocytochrome b2 or SEQ ID NO:7. However, there is no teaching regarding which amino acids can vary from any flavocytochrome b2, Kluyveromyces marxianus flavocytochrome b2 or SEQ ID NO:7 and result in polypeptide having flavocytochrome b2 activity. An important consideration is that structure is not necessarily a reliable indicator of function. In the instant case, there is no disclosure relating similarity of structure to conservation of function. Conservation of structure is not necessarily a surrogate for conservation of function. Since the claimed invention is that of a protein, and there is no disclosure of the domains responsible for flavocytochrome b2 activity, the absence of information may be persuasive that those of skill in the art would not take the disclosure as generic.
Fransceus (J Ind Microbiol Biotechnol. 2017 May;44(4-5):687-695. – form PTO-892) reviews protein engineering techniques, such as random mutagenesis and recombination, directed evolution and iterative or combinatory saturation “hotspots”. Fransceus states that “a recurring problem, however, is choosing which amino acid positions should be targeted. Answering this question is not an easy feat and requires substantial insight in the relationship between an enzyme’s sequence or structure and its properties.” Sanavia (Computational and Structural Biotechnology Journal, Volume 18, 2020, Pages 1968-1979. – form PTO-892) discloses challenges in the prediction of protein stability in the occurrence of multiple mutations. “Multiple-point mutations are common variations of the protein sequence that may be needed in protein engineering when a single-point mutation is not enough to yield the desired stability change. Dealing with multiple-site variations adds another level of complexity beyond the prediction of the effect of a single variant on protein stability, since it requires the learning of many types of combinatorial effects”.
Given this lack of description of the representative species encompassed by the genus of the claims, the specification fails to sufficiently describe the claimed invention in such full, clear, concise, and exact terms that a skilled artisan would recognize that applicants were in possession of the inventions of claims 1, 3, and 5.
Claim Rejections - 35 USC § 103
In the event the determination of the status of the application as subject to AIA 35 U.S.C. 102 and 103 (or as subject to pre-AIA 35 U.S.C. 102 and 103) is incorrect, any correction of the statutory basis (i.e., changing from AIA to pre-AIA ) for the rejection will not be considered a new ground of rejection if the prior art relied upon, and the rationale supporting the rejection, would be the same under either status.
The following is a quotation of 35 U.S.C. 103 which forms the basis for all obviousness rejections set forth in this Office action:
A patent for a claimed invention may not be obtained, notwithstanding that the claimed invention is not identically disclosed as set forth in section 102, if the differences between the claimed invention and the prior art are such that the claimed invention as a whole would have been obvious before the effective filing date of the claimed invention to a person having ordinary skill in the art to which the claimed invention pertains. Patentability shall not be negated by the manner in which the invention was made.
The factual inquiries for establishing a background for determining obviousness under 35 U.S.C. 103 are summarized as follows:
1. Determining the scope and contents of the prior art.
2. Ascertaining the differences between the prior art and the claims at issue.
3. Resolving the level of ordinary skill in the pertinent art.
4. Considering objective evidence present in the application indicating obviousness or nonobviousness.
This application currently names joint inventors. In considering patentability of the claims the examiner presumes that the subject matter of the various claims was commonly owned as of the effective filing date of the claimed invention(s) absent any evidence to the contrary. Applicant is advised of the obligation under 37 CFR 1.56 to point out the inventor and effective filing dates of each claim that was not commonly owned as of the effective filing date of the later invention in order for the examiner to consider the applicability of 35 U.S.C. 102(b)(2)(C) for any potential 35 U.S.C. 102(a)(2) prior art against the later invention.
Claim(s) 1, 3, and 5 is/are rejected under 35 U.S.C. 103 as being unpatentable over W0T5A6 (UnitProtKB/TrEMBL Database. February 26, 2020 – form PTO-892), Beasley (The signal that sorts yeast cytochrome b2 to the mitochondrial intermembrane space contains three distinct functional regions. EMBO J. 1993 Jun;12(6):2303-11 – from PTO-1449), Sun (Enhancing the Solubility of Recombinant Proteins in Escherichia coli by Using Hexahistidine-Tagged Maltose-Binding Protein as a Fusion Partner. In: Evans, Jr., T., Xu, MQ. (eds) Heterologous Gene Expression in E. coli. Methods in Molecular Biology, vol 705. Humana Press. (2011). – form PTO-892), and Smutok (A novel L-lactate-selective biosensor based on flavocytochrome b2 from methylotrophic yeast Hansenula polymorpha. Biosens Bioelectron. 2005 Jan 15;20(7):1285-90 – form PTO-892).
Regarding claim 5, W0T5A6 discloses a Kluyveromyces marxianus flavocytochrome b2 having 100% sequence identity to the flavocytochrome b2 of SEQ ID NO:7 of the instant application (pages 1-2 and see the sequence alignment below). W0T5A6 discloses that the amino acids 89-166 is the cytochrome heme binding domain and amino acids 198-565 is the FMN domain (page 2).
W0T5A6 does not teach an electrode comprising a truncated flavocytochrome b2 comprising a His tag, wherein the flavocytochrome b2 from the N-to C-terminus has viii) a methionine, ix. a tag sequence, x. an amino acid of a length of 0 of the native signal peptide, and xi. I/L/V-x-N/A/L motif of number 0.
Regarding claim 5, Beasley discloses that cytochrome b2 comprise an 80 residue presequence, which targets the cytochrome b2 from the cytoplasm into the mitochondrial membrane (abstract and page 2303).
Regarding claims 3 and 5, Sun discloses using a His tag-MBP as a fusion partner to enhance solubility of recombinant proteins and expression of recombinant proteins with the initial Met as the start codon (abstract, Figure 16.1, pages 259-260, and Sections 3.1.2 and 3.2).
Smutok discloses a sensor comprising an electrode comprising a flavocytochrome b2 to measure L-lactate (abstract).
Therefore, in combining the above references, it would have been obvious to one having ordinary skill in the art before the time the claimed invention was effectively filed to replace the presequence of amino acids 1-88 of the Kluyveromyces marxianus flavocytochrome b2 of W0T5A6 with an initial Met residue and the His tag-MBP, express and purify the flavocytochrome b2, and place the flavocytochrome b2 into an electrode. One of ordinary skill in the art at the time the invention was made would have been motivated to replace the presequence of the Kluyveromyces marxianus flavocytochrome b2 of W0T5A6 in order to enhance solubility of said flavocytochrome b2 for expression and purification. One of ordinary skill in the art at the time the invention was made would have been motivated to place the flavocytochrome b2 in a sensor comprising an electrode to measure L-lactate. One of ordinary skill in the art would have had a reasonable expectation of success since W0T5A6 discloses a Kluyveromyces marxianus flavocytochrome b, W0T5A6 and Beasley that flavocytochrome b has presequence, Sun discloses using a His-MBP as a fusion partner for enhanced solubility of a recombinant protein and expression and purification of the recombaint protein, and Smutok discloses using a sensor comprising an electrode comprising a flavocytochrome b to measure L-lactate.
Therefore, the above references render claims 1, 3, and 5 prima facie obvious
Conclusion
Claims 1-5 are pending.
Claims 1-5 are rejected.
Any inquiry concerning this communication or earlier communications from the examiner should be directed to YONG D PAK whose telephone number is (571)272-0935. The examiner can normally be reached M-Th: 5:30 am - 3:30 pm.
Examiner interviews are available via telephone, in-person, and video conferencing using a USPTO supplied web-based collaboration tool. To schedule an interview, applicant is encouraged to use the USPTO Automated Interview Request (AIR) at http://www.uspto.gov/interviewpractice.
If attempts to reach the examiner by telephone are unsuccessful, the examiner’s supervisor, Robert Mondesi can be reached on 408-918-7584. The fax phone number for the organization where this application or proceeding is assigned is 571-273-8300.
Information regarding the status of published or unpublished applications may be obtained from Patent Center. Unpublished application information in Patent Center is available to registered users. To file and manage patent submissions in Patent Center, visit: https://patentcenter.uspto.gov. Visit https://www.uspto.gov/patents/apply/patent-center for more information about Patent Center and https://www.uspto.gov/patents/docx for information about filing in DOCX format. For additional questions, contact the Electronic Business Center (EBC) at 866-217-9197 (toll-free). If you would like assistance from a USPTO Customer Service Representative, call 800-786-9199 (IN USA OR CANADA) or 571-272-1000.
/YONG D PAK/Primary Examiner, Art Unit 1652
Sequence alignment between the flavocytochrome b2 of SEQ ID NO:7 of the instant application (“Qy”) and the Kluyveromyces marxianus flavocytochrome b2 of W0T5A6 (“Db”)
W0T5A6_KLUMD
ID W0T5A6_KLUMD Unreviewed; 592 AA.
AC W0T5A6;
DT 19-MAR-2014, integrated into UniProtKB/TrEMBL.
DT 19-MAR-2014, sequence version 1.
DT 28-JAN-2026, entry version 53.
DE RecName: Full=L-lactate dehydrogenase (cytochrome) {ECO:0000256|ARBA:ARBA00068515};
DE EC=1.1.2.3 {ECO:0000256|ARBA:ARBA00066458};
DE AltName: Full=Cytochrome b2 {ECO:0000256|ARBA:ARBA00078774};
DE AltName: Full=Flavocytochrome b2 {ECO:0000256|ARBA:ARBA00075949};
DE AltName: Full=L-lactate ferricytochrome c oxidoreductase {ECO:0000256|ARBA:ARBA00078938};
GN Name=CYB2 {ECO:0000313|EMBL:BAO38243.1};
GN ORFNames=KLMA_10621 {ECO:0000313|EMBL:BAO38243.1};
OS Kluyveromyces marxianus (strain DMKU3-1042 / BCC 29191 / NBRC 104275)
OS (Yeast) (Candida kefyr).
OC Eukaryota; Fungi; Dikarya; Ascomycota; Saccharomycotina; Saccharomycetes;
OC Saccharomycetales; Saccharomycetaceae; Kluyveromyces.
OX NCBI_TaxID=1003335 {ECO:0000313|EMBL:BAO38243.1, ECO:0000313|Proteomes:UP000065495};
RN [1] {ECO:0000313|EMBL:BAO38243.1, ECO:0000313|Proteomes:UP000065495}
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=DMKU3-1042 / BCC 29191 / NBRC 104275
RC {ECO:0000313|Proteomes:UP000065495};
RX PubMed=25834639; DOI=10.1186/s13068-015-0227-x;
RA Lertwattanasakul N., Kosaka T., Hosoyama A., Suzuki Y., Rodrussamee N.,
RA Matsutani M., Murata M., Fujimoto N., Suprayogi, Tsuchikane K., Limtong S.,
RA Fujita N., Yamada M.;
RT "Genetic basis of the highly efficient yeast Kluyveromyces marxianus:
RT complete genome sequence and transcriptome analyses.";
RL Biotechnol. Biofuels 8:47-47(2015).
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(S)-lactate + 2 Fe(III)-[cytochrome c] = 2 Fe(II)-[cytochrome
CC c] + pyruvate + 2 H(+); Xref=Rhea:RHEA:19909, Rhea:RHEA-COMP:10350,
CC Rhea:RHEA-COMP:14399, ChEBI:CHEBI:15361, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:16651, ChEBI:CHEBI:29033, ChEBI:CHEBI:29034; EC=1.1.2.3;
CC Evidence={ECO:0000256|ARBA:ARBA00052399};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:19910;
CC Evidence={ECO:0000256|ARBA:ARBA00052399};
CC -!- COFACTOR:
CC Name=FMN; Xref=ChEBI:CHEBI:58210;
CC Evidence={ECO:0000256|ARBA:ARBA00001917};
CC -!- COFACTOR:
CC Name=heme b; Xref=ChEBI:CHEBI:60344;
CC Evidence={ECO:0000256|ARBA:ARBA00001970};
CC -!- SUBUNIT: Homotetramer. {ECO:0000256|ARBA:ARBA00011881}.
CC -!- SUBCELLULAR LOCATION: Mitochondrion intermembrane space
CC {ECO:0000256|ARBA:ARBA00004569}.
CC -!- SIMILARITY: In the C-terminal section; belongs to the FMN-dependent
CC alpha-hydroxy acid dehydrogenase family.
CC {ECO:0000256|ARBA:ARBA00061137}.
CC -!- SIMILARITY: In the N-terminal section; belongs to the cytochrome b5
CC family. {ECO:0000256|ARBA:ARBA00061589}.
CC ---------------------------------------------------------------------------
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DR EMBL; AP012213; BAO38243.1; -; Genomic_DNA.
DR RefSeq; XP_022674134.1; XM_022822298.1.
DR AlphaFoldDB; W0T5A6; -.
DR SMR; W0T5A6; -.
DR GeneID; 34714278; -.
DR KEGG; kmx:KLMA_10621; -.
DR VEuPathDB; FungiDB:KLMA_10621; -.
DR OrthoDB; 1925334at2759; -.
DR Proteomes; UP000065495; Chromosome 1.
DR GO; GO:0005758; C:mitochondrial intermembrane space; IEA:UniProtKB-SubCell.
DR GO; GO:0020037; F:heme binding; IEA:InterPro.
DR GO; GO:0004460; F:L-lactate dehydrogenase (cytochrome) activity; IEA:UniProtKB-EC.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0006089; P:lactate metabolic process; IEA:TreeGrafter.
DR CDD; cd02922; FCB2_FMN; 1.
DR FunFam; 3.10.120.10:FF:000009; Cytochrome b2, mitochondrial, putative; 1.
DR FunFam; 3.20.20.70:FF:000062; Cytochrome b2, mitochondrial, putative; 1.
DR Gene3D; 3.20.20.70; Aldolase class I; 1.
DR Gene3D; 3.10.120.10; Cytochrome b5-like heme/steroid binding domain; 1.
DR InterPro; IPR013785; Aldolase_TIM.
DR InterPro; IPR001199; Cyt_B5-like_heme/steroid-bd.
DR InterPro; IPR036400; Cyt_B5-like_heme/steroid_sf.
DR InterPro; IPR018506; Cyt_B5_heme-BS.
DR InterPro; IPR000262; FMN-dep_DH.
DR InterPro; IPR037396; FMN_HAD.
DR InterPro; IPR008259; FMN_hydac_DH_AS.
DR InterPro; IPR037458; L-MDH/L-LDH_FMN-bd.
DR PANTHER; PTHR10578:SF148; L-LACTATE DEHYDROGENASE (CYTOCHROME); 1.
DR PANTHER; PTHR10578; S -2-HYDROXY-ACID OXIDASE-RELATED; 1.
DR Pfam; PF00173; Cyt-b5; 1.
DR Pfam; PF01070; FMN_dh; 1.
DR SMART; SM01117; Cyt-b5; 1.
DR SUPFAM; SSF55856; Cytochrome b5-like heme/steroid binding domain; 1.
DR SUPFAM; SSF51395; FMN-linked oxidoreductases; 1.
DR PROSITE; PS00191; CYTOCHROME_B5_1; 1.
DR PROSITE; PS50255; CYTOCHROME_B5_2; 1.
DR PROSITE; PS00557; FMN_HYDROXY_ACID_DH_1; 1.
DR PROSITE; PS51349; FMN_HYDROXY_ACID_DH_2; 1.
PE 3: Inferred from homology;
KW Electron transport {ECO:0000256|ARBA:ARBA00022982};
KW Flavoprotein {ECO:0000256|ARBA:ARBA00022630};
KW FMN {ECO:0000256|ARBA:ARBA00022643}; Heme {ECO:0000256|ARBA:ARBA00022617};
KW Iron {ECO:0000256|ARBA:ARBA00023004};
KW Metal-binding {ECO:0000256|ARBA:ARBA00022723};
KW Mitochondrion {ECO:0000256|ARBA:ARBA00023128};
KW Oxidoreductase {ECO:0000256|ARBA:ARBA00023002};
KW Respiratory chain {ECO:0000256|ARBA:ARBA00022660};
KW Transit peptide {ECO:0000256|ARBA:ARBA00022946};
KW Transport {ECO:0000256|ARBA:ARBA00022448}.
FT DOMAIN 89..166
FT /note="Cytochrome b5 heme-binding"
FT /evidence="ECO:0000259|PROSITE:PS50255"
FT DOMAIN 198..564
FT /note="FMN hydroxy acid dehydrogenase"
FT /evidence="ECO:0000259|PROSITE:PS51349"
SQ SEQUENCE 592 AA; 64707 MW; C229AAC052607215 CRC64;
Query Match 100.0%; Score 3027; Length 592;
Best Local Similarity 100.0%;
Matches 592; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 MRSAARVINKSCSGSALSRRCLRKSSLSMSMRYLSTSNIGVRKGFNGQGKSSNKTMLFLA 60
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Db 1 MRSAARVINKSCSGSALSRRCLRKSSLSMSMRYLSTSNIGVRKGFNGQGKSSNKTMLFLA 60
Qy 61 AGASAVAGIGLLSQFSDSLQNATKEELNKPKVSPLEVAKHSSPDDCWVVIDGFVYNLTEF 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 61 AGASAVAGIGLLSQFSDSLQNATKEELNKPKVSPLEVAKHSSPDDCWVVIDGFVYNLTEF 120
Qy 121 ISAHPGGPAIIENNAGKDVTAIFGPIHAPDVIEKYIAPENRIGPLDGKMPDDLICAPLTP 180
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 121 ISAHPGGPAIIENNAGKDVTAIFGPIHAPDVIEKYIAPENRIGPLDGKMPDDLICAPLTP 180
Qy 181 GETPEDVARKEELRQNMPDLDSLVNIYDFEFLASQILTKQAWSYYSSAADDEVTHRENHA 240
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 181 GETPEDVARKEELRQNMPDLDSLVNIYDFEFLASQILTKQAWSYYSSAADDEVTHRENHA 240
Qy 241 AYHRIFFKPRILVNVKEVDTSTTMLGEKVGVPFYVSATALCKLGNPKEGEKDIARGCGES 300
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 241 AYHRIFFKPRILVNVKEVDTSTTMLGEKVGVPFYVSATALCKLGNPKEGEKDIARGCGES 300
Qy 301 DVKPIQMISTLASCSLQEIVEAAPSKDQIQWFQLYVNSDRKITEELIKNVEKLGLKAIFV 360
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 301 DVKPIQMISTLASCSLQEIVEAAPSKDQIQWFQLYVNSDRKITEELIKNVEKLGLKAIFV 360
Qy 361 TVDAPSLGNREKDAKVKFTNKDSSAKAMEKSNVKESKGASRALSTFIDPALCWDDIVTLK 420
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 361 TVDAPSLGNREKDAKVKFTNKDSSAKAMEKSNVKESKGASRALSTFIDPALCWDDIVTLK 420
Qy 421 SKTKLPIVIKGVQCVEDVLKAAEIGAAGVVLSNHGGRQLDFSRAPIEVLAETMPILKEKK 480
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 421 SKTKLPIVIKGVQCVEDVLKAAEIGAAGVVLSNHGGRQLDFSRAPIEVLAETMPILKEKK 480
Qy 481 LDDKIEIFIDGGVRRGTDILKALCLGAKGVGLGRPFLYANSCYGKEGVKKAIELLKDELE 540
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 481 LDDKIEIFIDGGVRRGTDILKALCLGAKGVGLGRPFLYANSCYGKEGVKKAIELLKDELE 540
Qy 541 MSMRLLGVTSIDQLSEKYLDLSTLHGRTVSVPRDNLYNGVYVPHEPTDFKEN 592
||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 541 MSMRLLGVTSIDQLSEKYLDLSTLHGRTVSVPRDNLYNGVYVPHEPTDFKEN 592
Prosecution Insights